Formulations of 4-(7-Hydroxy-2-isopropyl-4-oxo-4H-quinazolin-3-yl)-benzonitrile

ABSTRACT

The present invention provides formulations of 4-(7-hydroxy-2-isopropyl-4-oxo-4H-quinazolin-3-yl)-benzonitrile (compound I) and methods for treating ocular surface pain by administering such formulations. The present invention also provides methods for treating dry eye disease and ocular hyperemia by administering formulations of 4-(7-hydroxy-2-isopropyl-4-oxo-4H-quinazolin-3-yl)-benzonitrile.

REFERENCE TO RELATED APPLICATIONS

The application claims priority to U.S. application Ser. No. 16/789,976,filed Feb. 13, 2020, which claims benefit of U.S. ProvisionalApplication No. 62/806,705, filed Feb. 15, 2019, which is incorporatedherein in its entirety.

FIELD OF THE INVENTION

The present disclosure relates to formulations of4-(7-Hydroxy-2-isopropyl-4-oxo-4H-quinazolin-3-yl)-benzonitrile (FormulaI) and methods for treating ocular surface diseases using same as wellas methods for reducing ocular surface pain.

BACKGROUND OF THE INVENTION

The ocular surface, particularly the cornea, is densely innervated bysensory nerves. The activity of corneal nerves can be modified byinflammation caused by a number of factors, such as osmotic stress andtissue damage, as well as nerve injuries of the ocular surface. Ocularsurface symptoms are the alarm system to indicate an imbalanced ocularsurface homeostasis resulting in chronic ocular surface pain due tocontinuous stimuli causing stress and sensitization of the ocularsurface.

Patients suffering from ocular surface pain, particularly chronic ocularsurface pain have a significant decline in quality of life. In utilitystudies to date, the burden of severe chronic ocular surface pain hasbeen likened to moderate to severe angina, dialysis, or disabling hipfracture. Severe chronic ocular surface pain as also been associatedwith depression and suicidal ideation. In many patients, the ocularsurface pain remains unresolved despite treatment of the underlyingpathology (e.g., recent trauma or surgery, infection, or inflammation).Moreover, treatments that are used for short term management of ocularpain (e.g., nonsteroidal anti-inflammatory drugs, steroids, antibiotics)cannot be used for long term therapy. Thus, there is a long-felt andunmet need for safe, effective formulations for the symptomatictreatment of ocular surface pain when there are no other options toimprove patients' quality of life, or to supplement current treatments.

While topical administration of aqueous compositions is non-invasive andvery convenient, it remains a challenge to formulate hydrophobiccompounds into stable, aqueous formulations. Agglomeration can beparticularly troublesome for the hydrophobic ophthalmic drugs, which areparticularly prone to agglomeration within aqueous topical ophthalmiccompositions. Agglomeration may cause stability and potentially otherquality issues for the compositions, and may arise from otherinteractions of drugs and excipients.

In view of the above, it would be particularly desirable to provide anophthalmic composition, which can be dosed topically for the treatmentof ocular surface conditions, particular the treatment of ocular surfacepain.

SUMMARY OF THE INVENTION

In some embodiments, described herein is an aqueous formulation thatincludes:

4-(7-Hydroxy-2-isopropyl-4-oxo-4H-quinazolin-3-yl)-benzonitrile(compound I) or a salt, co-crystal, or polymorph thereof,

and one or more excipients selected from the group consisting of asurfactant, a suspending agent, a tonicity agent, a buffer, a salt, anda preservative.

In some embodiments, the4-(7-Hydroxy-2-isopropyl-4-oxo-4H-quinazolin-3-yl)-benzonitrile(compound I) or a salt, co-crystal, or polymorph thereof is present inthe formulation as a suspension. In alternative or additionalembodiments, the4-(7-Hydroxy-2-isopropyl-4-oxo-4H-quinazolin-3-yl)-benzonitrile(compound I) or a salt, co-crystal, or polymorph thereof is present inthe formulation in an amount of about 0.5% w/v to about 3.5% w/v.

In some embodiments, described herein is an aqueous formulation thatincludes:4-(7-Hydroxy-2-isopropyl-4-oxo-4H-quinazolin-3-yl)-benzonitrile(compound I) or a salt, co-crystal, or polymorph thereof, in an amountof about 0.5% w/v to about 3.5% w/v, present as a suspension in theformulation,

a surfactant,

a suspending agent,

and one or more excipients selected from the group consisting of atonicity agent, a buffer, a salt, and a preservative.

In some embodiments, the invention described herein is a formulationthat includes:

a suspension of4-(7-Hydroxy-2-isopropyl-4-oxo-4H-quinazolin-3-yl)-benzonitrile(compound I) or a salt, co-crystal, or polymorph thereof, in an amountof about 0.5% w/v to about 3.5% w/v,

a non-ionic surfactant;

a suspending agent;

a tonicity agent;

a buffer;

a salt; and

optionally, a preservative.

In some embodiments, the present disclosure relates to a formulation,comprising:

a suspension of4-(7-Hydroxy-2-isopropyl-4-oxo-4H-quinazolin-3-yl)-benzonitrile(compound I) or a salt, co-crystal, or polymorph thereof, in an amountof about 0.5% w/v to about 3.5% w/v,

a surfactant selected from the group consisting of non-ionic, anionic,cationic surfactants, and combinations thereof;

a suspending agent;

a tonicity agent;

a buffer;

optionally, a salt;

optionally, a preservative; and

water in quantity sufficient (qs) to 100%.

In some embodiments, the formulation includes a non-ionic surfactant. Insome embodiments of the formulations described herein, the non-ionicsurfactant is selected from the group consisting of a polysorbatesurfactant, a block copolymer of ethylene oxide, propylene oxidesurfactant, poloxamer, tyloxapol, and combinations thereof.

In some embodiments of the formulations described herein, the non-ionicsurfactant is tyloxapol, which present in an amount at least about0.001% w/v, at least about 0.01% w/v, at least about 0.02% w/v, leastabout 0.03% w/v, or at least about 0.04% w/v, and no more than about 1%w/v, no more than about 0.5% w/v, no more than about 0.3% w/v, or nomore than about 0.2% w/v, no more than about 0.1% w/v, or no more thanabout 0.08% w/v. In some embodiments, the tyloxapol is present in anamount of about 0.03% w/v to 0.08% w/v, or about 0.05% w/v.

In some embodiments of the formulations described herein, the non-ionicsurfactant is poloxamer in an amount of about 15 to about 20% w/v of theformulation.

In some embodiments of the formulations described herein, the suspendingagent is selected from the group consisting of carbomer, hydroxypropylmethyl cellulose (hypromellose), polyethylene glycol, and combinationsthereof. In some embodiments, the suspending agent is carbomer, presentin the formulation in an amount of at least about 0.05% w/v, at leastabout 0.1% w/v, or at least about 0.2% w/v, and no greater than about1.0% w/v, no greater than about 0.6% w/v, or no greater than about 0.5%.In some embodiments, the carbomer is present in the formulation in anamount of 0.1% w/v to about 0.3% w/v, or about 0.2% w/v.

In some embodiments of the formulations described herein, the suspendingagent is hydroxypropyl methyl cellulose present in the formulation in anamount of at least about 0.05% w/v, at least about 0.1% w/v, or at leastabout 0.25% w/v, and less than about 1.8% w/v, less than about 1.0% w/v,less than about 0.8% w/v, or less than about 0.6% w/v. In someembodiments, the suspending agent is a polyethylene glycol (PEG) havingmolecular weight of from about 200 to about 20,000 Da. In someembodiments, the suspending agent is PEG400, at a concentration of fromabout 4% w/v to about 9% w/v, about 5% w/v to about 8% w/v, or about 7%w/v, or PEG6000 at a concentration of from about 1% w/v to about 4% w/v,about 1% w/v to about 3% w/v, or about 2% w/v.

In some embodiments of the formulations described herein, the suspendingagent is substantially all carbomer homopolymer Type B.

In some embodiments of the formulations described herein, the tonicityagent is selected from the group consisting of polyols.

In some embodiments of the formulations described herein, the polyol isselected from the group selected from mannitol, glycerin, xylitol,sorbitol and propylene glycol, and combinations thereof. In someembodiments, the polyol is present in an amount from about 0.05% w/v toabout 10% w/v, from about 0.1% to about 8% w/v, from about 0.1% to about7% w/v, from about 0.1% to about 5% w/v. In particular embodiments, thepolyol is mannitol or glycerin, present in the formulation in an amountof from 0.1% w/v to about 5% w/v, or about 0.2% w/v, about 0.3% w/v,about 0.4% w/v, about 0.5% w/v, about 1% w/v, about 2% w/v, about 2.5%w/v, about 3.0% w/v, about 3.5% w/v, about 4.0% w/v, about 4.5% w/v, orabout 5% w/v.

In some embodiments of the formulations described herein, the buffer isselected from the group consisting of acetate, ascorbate, borate,hydrogen carbonate, carbonate, citrate, edetate (EDTA) gluconate,lactate, phosphate, propionate and TRIS (tromethamine). In particularembodiments, the buffer is phosphate or TRIS.

In some embodiments of the formulations described herein, the salt issodium chloride or potassium chloride.

In some embodiments of the formulations described herein, the suspendingagent is carbopol (carbomer homopolymer Type B) and amount of sodiumchloride is adjusted to an amount to provide a viscosity of theformulation of about 20 cP to about 200 cP, when using spindle CP-42 at60 rpm at about 25° C. In some embodiments, the sodium chloride ispresent in an amount from about 0.01% w/v to about 0.5% w/v, from about0.02% w/v to about 0.4% w/v, from about 0.03% w/v to about 0.3% w/v,from about 0.04% w/v to about 0.2% w/v, from about 0.05% w/v to about0.1% w/v, or about 0.05% w/v.

In some embodiments of the formulations described herein, the pH of theformulation is about 5.5 to about 8.0. In some embodiments, the pH ofthe formulation is from about 6.0 to about 8.0, about 6.0, or about 7.4.In some embodiments, the pH of the formulation is about 5.0 to about8.0, about 5.5 to about 7.5, about 5.0 to about 7.4, about 5.5 to about7.4, about 6.0 to about 8.0, about 6.5 to about 8.0, about 6.0 to about7.4, or about 6.5 to about 7.4.

In some embodiments the formulations described herein further include anadditional agent selected from the group consisting of cyclodextrins inan amount of at least about 1.5 w/v %, at least about 3.0 w/v %, atleast about 3.5 w/v % or at least about 4.5 w/v, but no greater thanabout 10.0 w/v %, no greater than about 8.0% w/v, no greater than about6.5 w/v %, or no greater than about 5.5 w/v. In some embodiments, thecyclodextrin is hydroxypropyl β-cyclodextrin or sulfoalkyletherβ-cyclodextrin in an amount of about 5% w/v of the formulation.

In some embodiments, the present disclosure is related to a formulation,comprising:

the compound I or a salt, co-crystal, or polymorph thereof, is in anamount of about 0.5% w/v to about 2.5% w/v,

the a non-ionic surfactant is tyloxapol, poloxamer, or combinationsthereof, in an amount of from about 0.01 to 0.2% w/v;

the a suspending agent is hydroxypropyl methyl cellulose, polyethyleneglycol or carbomer homopolymer Type B;

the a tonicity agent is at least one polyol in an amount of from about0.05% w/v to about 10% w/v;

the buffer is edetate, phosphate, borate, or combinations thereof

a salt; and

water qs to 100%; and

the pH is in the range of from about 5.5 to about 8.0.

In some embodiments, the present disclosure is related to a formulation,comprising:

compound I or a salt, co-crystal, or polymorph thereof, is present in anamount of about 0.5% w/v, about 1.0% w/v, about 1.5% w/v, about 2.0%w/v, or about 2.5% w/v,

a non-ionic surfactant, which is tyloxapol in an amount of about 0.04w/v to about 0.06% w/v, poloxamer in an amount of about 0.005-0.12% w/v,or combinations thereof;

a suspending agent, which is hydroxypropyl methyl cellulose in an amountof from about 0.1% w/v to about 0.8 w/v %, polyethylene glycol in anamount of from about 2% w/v to about 8% w/v, carbomer homopolymer Type Bin an amount from about 0.05% w/v to about 0.5% w/v, or combinationsthereof;

a tonicity agent which is mannitol or glycerin in an amount of fromabout 0.1% w/v to about 5% w/v;

a buffer which is edetate, phosphate, borate, tromethamine, orcombinations thereof;

sodium chloride in an amount of from 0.01% w/v to about 1% w/v; and

water qs to 100%; and

having a pH is in the range of from about 5.5 to about 8.0.

In some embodiments, the present disclosure is related to a formulation,comprising:

a suspension of4-(7-Hydroxy-2-isopropyl-4-oxo-4H-quinazolin-3-yl)-benzonitrile(compound I) or a salt, co-crystal, or polymorph thereof, in an amountof about 0.5% w/v, about 1.0% w/v, about 1.5% w/v, about 2.0% w/v, orabout 2.5% w/v,

tyloxapol in an amount of about 0.04 w/v to about 0.06% w/v;

carbomer homopolymer Type B in an amount from about 0.05% w/v to about0.4% w/v;

glycerin in an amount of from about 0.5% w/v to about 5% w/v;

a buffer selected from the group consisting of edetate, phosphate,borate, tomethamine, and combinations thereof;

sodium chloride in an amount of from 0.01% w/v to about 1% w/v; and

water qs to 100%;

wherein the formulation has a pH in the range of from about 5.5 to about8.0.

In some embodiments of the formulations described herein, compound I isin polymorphic form B.

In some embodiments, the present disclosure is related to a formulation,comprising:

a suspension of polymorphic form B of4-(7-Hydroxy-2-isopropyl-4-oxo-4H-quinazolin-3-yl)-benzonitrile(compound I), in an amount of about 0.5% w/v, about 1.0% w/v, about 1.5%w/v, about 2.0% w/v, or about 2.5% w/v,

tyloxapol in an amount of about 0.04 w/v to about 0.06% w/v;

carbomer homopolymer Type B in an amount from about 0.05% w/v to about0.4% w/v;

glycerin in an amount of from about 0.5% w/v to about 5% w/v;

a buffer selected from edetate, phosphate, borate, tomethamine, orcombinations thereof;

sodium chloride in an amount of from 0.01% w/v to about 1% w/v; and

water qs to 100%;

wherein the formulation has a pH in the range of from about 5.5 to about8.0.

In some embodiments, the formulation comprises:

compound I in an amount of about 0.5% w/v, about 1.0% w/v, about 1.5%w/v, about 2.0% w/v, or about 2.5% w/v,

about 0.05% w/v of tyloxapol;

about 0.2% w/v of carbomer homopolymer Type B;

about 2.0% of glycerin;

a tromethamine buffer; and

hydrochloric acid to adjust pH to about 6.4 to about 8.4;

about 0.05% w/v of sodium chloride; and

water qs to 100%;

wherein the formulation does not include a preservative.

In some embodiments of the formulations described herein, thepolymorphic form B of compound I is characterized by an X-raydiffraction pattern having three or more peaks at 2θ values selectedfrom 9.3, 10.6 and 14.4.+−.0.2° 2θ.

In some embodiments, the formulations described herein have a viscosityof about 20 cP to about 200 cP.

In some embodiments, the formulations described herein have anosmolality of about 200 to about 450 milliosmoles per kilogram(mOsm/kg).

In some embodiments of the formulations described herein, the D₉₀ ofcompound I (diameter at which 90% of compound I is comprised of smallerparticles) is below about 10 μm, below about 8 μm, below about 6 μm,below about 4 μm, below about 3 μm, or about 2 μm. In some embodiments,the D₅₀ of compound I in the formulation (diameter at which 50% ofcompound I is comprised of smaller particles) is below about 10 μm,below about 8 μm, below about 6 μm, below about 4 μm, below about 3 μm,below about 2 μm, or about 1 μm. In some embodiments, the D₁₀ ofcompound I in the formulation (diameter at which 10% of compound I iscomprised of smaller particles) is below about 5 μm, below about 4 μm,below about 3 μm, below about 2 μm, below about 1 μm, or about 0.3 μm.

In some embodiments of the formulations described herein the formulationexhibits settling of less than about 10%, less than about 8%, less thanabout 7%, less than about 6%, less than about 5%, less than about 4%,less than about 3%, or less than about 2% after storage at roomtemperature for six months.

In some embodiments of the formulations described herein, the amount ofcompound I in the formulation is at least 90% of the initial amountafter about 6 months, after about 8 months, about 10 months, about 12months, about 15 months, or about 18 months of storage underrefrigeration.

In some embodiments of the formulations described herein, the amount ofcompound I in the formulation at least about 91%, at least about 92%, atleast about 93%, at least about 94%, at least about 95%, at least 96%,at least about 97% or at least about 98% of the initial amount afterabout 18 months of storage under refrigeration.

In some embodiments of the formulations described herein, theformulation comprises no more than about 10% of a degradation productafter 6 months under refrigeration, wherein the degradation product hasa relative retention time of 1.23, compared to compound I, when analyzedby HPMC using a gradient 0.1% trifluoroacetic acid (TFA)water/acetonitrile mobile phase.

In some embodiments of the formulations described herein, no more thanabout 10% of compound I in the formulation degrades upon storage for 12weeks at 40° C.

In some embodiments, the present disclosure is related to a method ofmaking a formulation, comprising

mixing an amount of4-(7-Hydroxy-2-isopropyl-4-oxo-4H-quinazolin-3-yl)-benzonitrile(compound I) or a salt, co-crystal, or polymorph thereof,

a non-ionic surfactant;

a suspending agent;

a tonicity agent;

a buffer;

a salt;

optionally, a preservative; and

water qs to 100%, and

adjusting the pH to a range of from about 5.5 to about 8.0.

In some embodiments, the method of making the formulations disclosedherein include the addition of compound I as a stock suspension. In someembodiments, the stock suspension is milled to achieve a desiredparticle size of compound I. In some embodiments, the D₉₀ of compound Iin the formulation (diameter at which 90% of compound I is comprised ofsmaller particles) is below about 10 μm, below about 8 μm, below about 6μm, below about 4 μm, below about 3 μm, or about 2 μm. In someembodiments, the D₅₀ of compound I in the formulation (diameter at which50% of compound I is comprised of smaller particles) is below about 10μm, below about 8 μm, below about 6 μm, below about 4 μm, below about 3μm, below about 2 μm, or about 1 μm. In some embodiments, the D₁₀ ofcompound I in the formulation (diameter at which 10% of compound I iscomprised of smaller particles) is below about 5 μm, below about 4 μm,below about 3 μm, below about 2 μm, below about 1 μm, or about 0.3 μm.

In one embodiment, the present disclosure provides a method of treatingocular surface pain in a subject in need thereof, comprising ocularlyadministering an effective amount of4-(7-Hydroxy-2-isopropyl-4-oxo-4H-quinazolin-3-yl)-benzonitrile (FormulaI) having structure:

or a salt, solvate, polymorph, or co-crystal thereof to the subject.

In some embodiments, the ocular surface pain is episodic or acute pain.In some embodiments, the ocular surface pain is chronic ocular surfacepain (COSP), lasting for at least about three months.

In some embodiments, the compound of formula I is administered to thecornea of the subject. In some embodiments, compound of formula I isadministered to the subject at a concentration of about 0.5% w/v toabout 3.5% w/v, about 0.5% w/v to about 2.5% w/v, or about 0.5% w/v toabout 1.5 w/v, about 0.5% to about 3.0% w/v, about 1.0% to about 2.5%w/v, about 1.5% to about 3.0% w/v, or about 0.5% to about 2.5% w/v. Inparticular embodiments, the compound of formula I is administered at aconcentration of about 0.5% w/v, about 1.0% w/v, about 1.5% w/v, about2.0% w/v, about 2.5% w/v, about 3.0% w/v, or about 3.5% w/v.

In some embodiments of the methods to treat ocular surface paindescribed herein, the COSP is associated with dry eye disease. In someembodiments, the administration results in a decrease in the symptoms ofdry eye disease. In particular embodiments, the administration resultsin a decrease in the ocular pain associated with dry eye disease. Insome embodiments, the administration results in reduced incidence of atleast about 10% in one or more of ocular dryness, ocular discomfort,ocular hyperemia, ocular burning or stinging, grittiness or foreign bodysensation, or photophobia.

In some embodiments of the methods to treat ocular surface paindescribed herein, the subject suffers from one or more of dry eyedisease, Sjogren's Syndrome, conjunctivitis (includingkeratoconjuctivitis, vernal keratoconjunctivitis, allergicconjunctivitis), Map-Dot-Fingerprint Dystrophy, acanthamoeba,fibromyalgia, Meibomian gland dysfunction, thyroid eye disease, rosacea,ptosis, keratoconus, ocular pain syndrome, Steven-Johnson's syndrome,corneal epitheliopathies, corneal neuropathies (including LASIK inducedcorneal neuropathies), corneal dystrophies (including recurrent cornealdystrophies), epithelial basement membrane dystrophy, corneal erosionsor abrasions (including recurrent corneal erosions or abrasions), ocularsurface diseases, blepharitis, graft vs host disease, meibomitis,glaucoma, conjunctivochalasis, keratopathis (including herpetickeratopathy, filamentary keratopathy, band or bullous keratopathy,exposure keratopathy), keratitis (including herpes simplex viruskeratitis), iritis, episclentis, corneal surgery, multiple sclerosis,trichiasis, pterygium, neuralgia, xerophthalmia, patients recoveringfrom neurotrophic keratitis, or ocular pain persisting for at leastthree months after photorefractive keratectomy (PRK) surgery orlaser-assisted in situ keratomileusis (LASIK) surgery.

In some embodiments, the methods to treat ocular surface pain describedherein include administering an additional therapeutic agent to thesubject.

In some embodiments of the methods to treat ocular surface paindescribed herein, the administration of compound I to the subjectresults in a reduction in a pain score of at least about 3, at leastabout 4, at least about 5, at least about 6, at least about 7, at leastabout 8, at least about 9 or at least about 10, compared to a placebo,when measured on a visual analog scale (VAS). In further embodiments,the administration of compound I to the subject results in a reductionin the subject's pain score of at least about 6, at least about 7, atleast about 8, at least about 9 or at least about 10, compared to aplacebo, when measured on the VAS.

In some embodiments of the methods to treat ocular surface paindescribed herein, the administration of compound I to the subjectresults in a reduction in the subject's pain of at least about 10%, atleast about 15%, at least about 20%, or at least about 25%, compared toa placebo. In some embodiments, the reduction in the pain score arisesfrom the difference in pain scores prior to and after administration ofcompound I to the subject. In some embodiments of the methods describedherein, the administration of compound I to the subject the reduction inpain score occurs within about half hour after administration ofcompound I to the subject.

In some embodiments of the methods to treat ocular surface paindescribed herein, the administration of compound I to the subjectresults in a reduction in hyperemia in the subject of at least about 1,at least about 2, at least about 3, at least about 4, or at least about5, on the McMonnies scale.

In some embodiments of the methods to treat ocular surface paindescribed herein, the administration of compound I to the subjectresults in a reduction in pain score within about half hour, withinabout 1 hour, within about 2 hours, or within about 4 hours afteradministration of compound I to the subject.

In some embodiments of the methods described herein, the administrationof compound I to the subject the administration results in a reductionin hyperemia in the subject of at least about 1, at least about 2, atleast about 3, at least about 4, or at least about 5, on the McMonniesscale.

In some embodiments of the methods described herein, the administrationof compound I to the subject the administration does not result in achange in one or more of best corrected visual acuity, intraocularpressure, slit-lamp biomicroscopy, dilated eye exam, blink rate, or tearproduction, compared to a placebo.

In some embodiments of the methods described herein, the compound offormula I is administered in the form of a formulation described herein.In some embodiments of the methods described herein, the formulation isadministered for at least about one, about two, or about three months.In some embodiments, the formulation is administered one to four timesdaily.

In some embodiments, the disclosure is related to a formulation asdescribed herein for use in the treatment of ocular surface pain. Insome embodiments, the ocular surface pain is acute or episodic ocularsurface pain. In some embodiments, the ocular surface pain is chronicocular surface pain lasting for at least 1 month, at least 2 months, orat least 3 months.

In some embodiments, the present disclosure relates to a method ofreducing ocular surface pain in a subject in need thereof, comprisingocularly administering4-(7-Hydroxy-2-isopropyl-4-oxo-4H-quinazolin-3-yl)-benzonitrile (FormulaI) having structure:

or a salt, solvate, polymorph, or co-crystal thereof to the subject.

In some embodiments, the ocular surface pain is acute or episodic ocularsurface pain. In some embodiments, the ocular surface pain is chronicocular surface pain lasting for at least 1 month, at least 2 months, orat least 3 months. In some embodiments, the COSP is associated with dryeye disease.

In some embodiments of the methods to reduce ocular pain describedherein, the COSP is associated with dry eye disease. In some embodimentsof the methods to reduce ocular surface pain described herein, theadministration of compound I to the subject results in a decrease in thesymptoms of dry eye disease.

In some embodiments of the methods to reduce ocular surface paindescribed herein, the administration of compound I to the subjectresults in a decrease in the ocular pain associated with dry eyedisease. In some embodiments of the methods described herein, theadministration of compound I to the subject results in reduced incidenceof at least about 10% in one or more of ocular dryness, oculardiscomfort, ocular hyperemia, ocular burning or stinging, grittiness orforeign body sensation, or photophobia.

In some embodiments, the subject in need of reducing ocular surface painsuffers from one or more of dry eye disease, Sjogren's Syndrome,conjunctivitis (including keratoconjuctivitis, vernalkeratoconjunctivitis, allergic conjunctivitis), Map-Dot-FingerprintDystrophy, acanthamoeba, fibromyalgia, Meibomian gland dysfunction,thyroid eye disease, rosacea, ptosis, keratoconus, ocular pain syndrome,Steven-Johnson's syndrome, corneal epitheliopathies, cornealneuropathies (including LASIK induced corneal neuropathies), cornealdystrophies (including recurrent corneal dystrophies), epithelialbasement membrane dystrophy, corneal erosions or abrasions (includingrecurrent corneal erosions or abrasions), ocular surface diseases,blepharitis, graft vs host disease, meibomitis, glaucoma,conjunctivochalasis, keratopathis (including herpetic keratopathy,filamentary keratopathy, band or bullous keratopathy, exposurekeratopathy), keratitis (including herpes simplex virus keratitis),iritis, episclentis, corneal surgery, multiple sclerosis, trichiasis,pterygium, neuralgia, xerophthalmia, or patients recovering fromneurotrophic keratitis. In some embodiments, the subject suffers fromocular pain persisting for at least three months after photorefractivekeratectomy (PRK) surgery or laser-assisted in situ keratomileusis(LASIK) surgery.

In some embodiments, the methods to reduce ocular surface pain describedherein include administering an additional therapeutic agent to thesubject.

In some embodiments of the methods to reduce ocular surface paindescribed herein, the administration of compound I to the subjectresults in a reduction in the subject's pain score of at least about 3as compared to a pain score prior to administration of the compound,when measured on a visual analog scale (VAS). In some embodiments of themethods described herein, the administration of compound I to thesubject results in a reduction in a pain score of at least about 3, atleast about 4, at least about 5, at least about 6, at least about 7, atleast about 8, at least about 9 or at least about 10, compared to aplacebo, when measured on a visual analog scale (VAS). In someembodiments of the methods described herein, the administration ofcompound I to the subject results in a reduction in the subject's painscore of at least about 6, at least about 7, at least about 8, at leastabout 9 or at least about 10, compared to a placebo, when measured onthe VAS. In other embodiments, the reduction in pain score occurs afterabout 7 days of administration of compound I to the subject. In someembodiments, the reduction in pain score occurs after about 14 days ofadministration of compound I to the subject.

In some embodiments of the methods to reduce ocular surface paindescribed herein, the administration of compound I to the subjectresults in a reduction in the subject's pain of at least about 10%, atleast about 15%, at least about 20%, or at least about 25%, compared toa placebo. In some embodiments of the methods described herein, thereduction in the pain score arises from the difference in pain scoresprior to and after administration of compound I to the subject. In otherembodiments, the reduction in pain score occurs after about 7 days ofadministration of compound I to the subject. In some embodiments, thereduction in pain score occurs after about 14 days of administration ofcompound I to the subject.

In some embodiments of the methods to reduce ocular surface paindescribed herein, the administration of compound I to the subjectresults in a reduction in hyperemia in the subject of at least about 1,at least about 2, at least about 3, at least about 4, or at least about5, on the McMonnies scale.

In some embodiments of the methods described herein, the compound offormula I is administered as a formulation as described herein.

In some embodiments, the present disclosure relates to a method ofreducing ocular hyperemia in a subject in need thereof, comprisingocularly administering 4-(7-hydroxyisopropyl-4-oxo-4H-quinazolin-3-yl)-benzonitrile (Formula I) havingstructure:

or a salt, solvate, polymorph, or co-crystal thereof to the subject;wherein the compound of formula I is administered at a concentration ofabout 0.5% w/v to about 3.5% w/v, about 0.5% w/v to about 2.5% w/v, orabout 0.5% w/v to about 1.5 w/v, about 0.5% to about 3.0% w/v, about1.0% to about 2.5% w/v, about 1.5% to about 3.0% w/v, or about 0.5% toabout 2.5% w/v.

In some embodiments of the methods described herein, the reduction inocular hyperemia is at least about 1, at least about 2, at least about3, at least about 4, or at least about 5, on the McMonnies scale.

In some embodiments, the administration of compound I results in reducedincidence of at least about 10% in one or more of ocular dryness, oculardiscomfort, ocular burning or stinging, grittiness or foreign bodysensation, or photophobia.

In some embodiments of the methods described herein, the subject suffersfrom one or more of dry eye disease, Sjogren's Syndrome, conjunctivitis(including keratoconjuctivitis, vernal keratoconjunctivitis, allergicconjunctivitis), Map-Dot-Fingerprint Dystrophy, acanthamoeba,fibromyalgia, Meibomian gland dysfunction, thyroid eye disease, rosacea,ptosis, keratoconus, ocular pain syndrome, Steven-Johnson's syndrome,corneal epitheliopathies, corneal neuropathies (including LASIK inducedcorneal neuropathies), corneal dystrophies (including recurrent cornealdystrophies), epithelial basement membrane dystrophy, corneal erosionsor abrasions (including recurrent corneal erosions or abrasions), ocularsurface diseases, blepharitis, graft vs host disease, meibomitis,glaucoma, conjunctivochalasis, keratopathis (including herpetickeratopathy, filamentary keratopathy, band or bullous keratopathy,exposure keratopathy), keratitis (including herpes simplex viruskeratitis), iritis, episclentis, corneal surgery, multiple sclerosis,trichiasis, pterygium, neuralgia, xerophthalmia, patients recoveringfrom neurotrophic keratitis, or ocular pain persisting for at leastthree months after photorefractive keratectomy (PRK) surgery orlaser-assisted in situ keratomileusis (LASIK) surgery.

Some embodiments of the methods described herein to reduce ocularhyperemia further include administering an additional therapeutic agentto the subject.

In some embodiments of the methods described herein, the compound offormula I is administered as a formulation as described herein.

In some embodiments, the present disclosure relates to a method oftreating or reducing ocular surface pain in a subject in need thereof,comprising administering to the subject a formulation comprising acompound 4-(7-hydroxy-2-isopropyl-4-oxo-4H-quinazolin-3-yl)-benzonitrile(compound I) having structure:

wherein the formulation results in a rabbit corneal Cmax compound I ofabout 1.5 to about 3 times conjunctival Cmax, wherein Cmax is themaximum concentration of compound I in the specified tissue afteradministration of a single dose. In some embodiments, the compound I isadministered as a formulation as described herein.

In some embodiments, the present disclosure relates to a method oftreating or reducing ocular surface pain in a subject in need thereof,comprising administering to the subject a formulation comprising acompound 4-(7-hydroxy-2-isopropyl-4-oxo-4H-quinazolin-3-yl)-benzonitrile(compound I) having structure:

wherein the formulation results in a Cmax of compound I in a rabbitcornea of about 500 times the Cmax of compound I in plasma, wherein Cmaxis the maximum concentration of compound I in the specified tissue afteradministration of a single dose. In some embodiments, the compound I isadministered as a formulation as described herein.

Specific preferred embodiments of the invention will become evident fromthe following more detailed description of certain preferred embodimentsand the claims.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1A shows the percent of compound I in the exploratory formulationsdescribed in Table 2 through 12 weeks at room temperature. In FIG. 1A,the formulation with percent of compound I at about 70% of original at12 weeks is the formulation of compound I as a solution. FIG. 1B showsthe percent of compound I in the exploratory formulations after 12 weeksat 40° C. In FIG. 1B, the formulation with percent of compound I atabout 20% of original at 12 weeks is the formulation of compound I as asolution.

FIG. 2 shows the X-ray Powder Diffraction Patterns of compound IRecovered from 12-Week Stability Samples versus Control.

FIG. 3 shows the model estimated mean (+/−SE) of visual analog scale(VAS) pain assessment over time (Primary PD analysis set).

FIG. 4 shows a histogram of oral rescue medication use incidence (numberof patients who did not use oral rescue medication) (Secondary PDanalysis set). For each of the time periods, the right bar representssubjects who were administered compound I, while the left bar representssubjects who were administered vehicle. The percent of patients who didnot use oral rescue medication (ORM) is presented in the y-axis. n/Nrepresents the count/total number of patients at each treatment.

FIG. 5A-5C provides the arithmetic mean (+/−SD) of ocular painassessment survey (OPAS) over time for questions 4, 5, and 6 (SecondaryPD analysis set). FIG. 5A provides the results from question 4: 24 hrEye Pain When Most Painful (pain level), FIG. 5B provides results fromQuestion 5: 24 hr Eye Pain When Least Painful (pain level), and FIG. 5Cprovides results from Question 6 24 hr Eye Pain On Average (pain level).For each of FIGS. 5A, 5B, and 5C, the dashed line with “x” representsvehicle, and the solid line with circles (o) represents compound I.

FIG. 6A-6D provides the arithmetic mean (+/−SD) of ocular painassessment survey (OPAS) over time for questions 22, 23, 24 and 25(Secondary PD analysis set). FIG. 6A provides results from Question 22:How Often Eye Pain With Redness (%). FIG. 6B provides results fromQuestion 23: How Often Eye Pain With Burning (%). FIG. 6C providesresults from Question 24: How Often Eye Pain With Sensitivity (%). FIG.6D provides results from Question 25: How Often Eye Pain With Tearing(%). For each of FIGS. 6A, 6B, 6C, and 6D, the dashed line with “x”represents vehicle, and the solid line with circles (o) representscompound I.

FIG. 7A provides the arithmetic mean (SD) plasma concentrations ofcompound I following topical ocular administration of 2.5% compound I(PK analysis set) on Day 1.

FIG. 7B provides the arithmetic mean (SD) plasma concentrations ofcompound I following topical ocular administration of 2.5% compound I(PK analysis set) on Day 4.

FIG. 8A provides a bar chart of ocular hyperemia over time for compoundI, FIG. 8B provides a bar chart of ocular hyperemia over time forVehicle. The quadrants are S: Superior, N: Nasal, I: Inferior, and T:Temporal.

FIG. 9 provides the arithmetic mean (+/−SD) of epithelial defect sizeover time (Safety analysis set) Parameter (unit): Epithelial Wound Size(mm), Subcategory: Area (mm2). The dashed line with “x” representsvehicle, and the solid line with circles (o) represents compound I.

DETAILED DESCRIPTION

“TRPV1 receptor” refers to the Transient Receptor Potential Vanilloid 1that has been characterized through molecular cloning and pharmacology.See e.g., Caterina M J, et al., Nature 1997; 389:816-824. TRPV1 receptoractivity is measured as described in WO2005/120510, hereby incorporatedby reference in its entirety.

The language “effective amount” of the compounds described herein,refers to that amount of a therapeutic compound necessary or sufficientto perform its intended function within a mammal. An effective amount ofthe therapeutic compound can vary according to factors such as theamount of the causative agent already present in the mammal, the age,sex, and weight of the mammal, and the ability of the therapeuticcompounds of the present disclosure to treat the ocular surface disorderand/or symptoms thereof in the mammal.

The phrase “ophthalmically compatible” refers to formulations, polymersand other materials and/or dosage forms which are suitable for use incontact with the ocular tissues of human beings and animals withoutexcessive toxicity, irritation, allergic response, or other problem orcomplication, commensurate with a reasonable benefit/risk ratio.

As used herein, the term “treat”, “treating” or “treatment” inconnection to a disease or disorder refers in some embodiments, toameliorating the disease or disorder (i.e., slowing or arresting orreducing the development of the disease or at least one of the clinicalsymptoms thereof). In another embodiment “treat”, “treating” or“treatment” refers to alleviating or ameliorating at least one physicalparameter including those which may not be discernible by the patient.In yet another embodiment, “treat”, “treating” or “treatment” refers tomodulating the disease or disorder, either physically, (e.g.,stabilization of a discernible symptom), physiologically, (e.g.,stabilization of a physical parameter), or both. In yet anotherembodiment, “treat”, “treating” or “treatment” refers to preventing ordelaying the onset or development or progression of the disease ordisorder or symptom thereof.

As used herein, the term “subject” or “patient” refers to human andnon-human mammals, including but, not limited to, primates, rabbits,pigs, horses, dogs, cats, sheep, and cows. In particular embodiments, asubject or patient is a human. In some embodiments, the term “patient”or “subject” refers to a human being who is diseased with the condition(i.e., disease or disorder) described herein and who would benefit fromthe treatment. As used herein, a subject is “in need of” a treatment ifsuch subject (patient) would benefit biologically, medically or inquality of life from such treatment. In particular embodiments, thesubject is an adult human of at least about 18 years of age. In someembodiments, the subject is an adult human from about 18 years of age toabout 75 years of age. In some embodiments, the subject is a child of upto about 18 years of age.

As used herein, “ocular surface” refers to the outer surface of the eye,which anatomically comprises the cornea (with epithelium, bowman layer,stroma, descement membrane, endothelium), conjunctiva, cul de sac, andthe corneo-scleral junction, i.e., limbus.

As used herein, ocular administration includes administration to allparts of the eye including all parts of the ocular surface such as thecornea, conjunctiva, the cul de sac and the corneo-scleral junction,i.e., limbus.

As used herein, “pain” refers to constant or intermittent sensation ofactual pain described as but not limited to stabbing, dull, sharp, orache. Pain may also refer to similar related descriptors such as but notlimited to burning, stinging, grittiness, foreign body sensation,dryness, sandy, tired, itchy, irritated, sensitivity to light.

As used herein, “ocular surface pain” refers to pain on the surface ofthe eye, e.g., cornea. Ocular pain may be nociceptic pain, which isgenerally caused by external physical or chemical damaging stimuli suchas corneal surgery, inflammation, or other damage to the cornealsurface. Ocular pain may also result from neuropathic pain, which mayoccur due to direct damage to the neurons of the body, resulting inmessages of pain being sent to the central nervous system and brainregardless of the presence of noxious stimuli. As used herein “ocularsurface pain” includes both nociceptic pain and neuropathic pain.

As used herein, the term “visual analog scale” (VAS) is a measure ofpain intensity where a subject typically marks a place on a scale thataligns with their level of pain. The pain is marked in a range of “nopain” (score of 0) and “pain as bad as it could be” or “worst imaginablepain” (score of 100). See e.g., Hawker, et al., Arthritis Care &Research 63(11), pp. S240-S252 (November 2011). There are several otherwell-designed pain scales that may be used to help assess the extent ofpain. The numerical rating scale (NRS) is often used, in which subjectsuse numbers to rate pain. The number scale may be from 1-10, or 1-100.The Wong-Baker FACES Pain Scale combines pictures and numbers for painratings. It can be used in children over the age of 3 and in adults. Sixfaces depict different expressions, ranging from happy to extremelyupset. Each is assigned a numerical rating between 0 (smiling) and 10(crying). The Verbal Pain Intensity Scale uses wordings on a scale torate pain intensity: No Pain/Mild Pain/Moderate Pain/Severe Pain VerySevere Pain/Worst Possible Pain.

The Eye Sensation Scale is a specific pain scale was developed tomeasure ophthalmic pain severity. See Caudle L. E. et al., Optom VisSci. 2007 August; 84(8):752-62. In this scale, pain, discomfort or lightsensitivity is typically measured by 5 category labels of “extreme,”“severe,” “moderate,” “mild,” or “none.”

The Ocular Pain Assessment Survey (OPAS) is a quantitative,multidimensional questionnaire, specifically designed for assessment ofcorneal and ocular surface pain and Quality of Life (QoL) changes. TheOPAS assesses pain intensity, frequency of eye and non-eye pain, QoLchanges, aggravating factors, associated factors, and symptomatic reliefquantitative, allowing for monitoring of treatment responses. See Qaziet al., Ophthalmology July 123(7):1458-1468 (2016).

As used herein, the term “Visual Tasking Questionnaire” refers to aquestionnaire that asks the subject to subjectively rate how muchdifficulty they have conducting certain activities that require a fixedor prolonged stare that may exacerbate ocular pain. The questionnairealso asks about coping mechanisms associated with the difficulties theyexperience during visual tasking activities.

As used herein, ocular hyperemia refers to redness of the ocularsurface. Ocular hyperemia may be a clinical marker for inflammationand/or ocular irritation. Ocular hyperemia may be measured using theMcMonnies scale, at values from 0 to 5, based on standard photographs.

As used herein, “placebo” refers to an ophthalmic formulation thatincludes all the components of the administered drug composition withoutthe drug.

As used herein, the term “about” refers to a range of values+10% of aspecified value.

As used herein, “Compound of formula I,” “Compound I,” “Formula I,” and“compound I” are used interchangeably and mean a compound that has thename 4-(7-hydroxy-2-isopropyl-4-oxo-4H-quinazolin-3-yl)-benzonitrile,the structure shown below, and can be synthesized using procedures knownin the art and described in WO2005/120510 and U.S. Pat. No. 8,349,852(“Quinazolinone derivatives useful as vanilloid antagonists”) to Chen etal., both of which are hereby incorporated by reference in theirentireties.

Compound I may be used in amorphous or crystalline forms. Additionallyor alternatively, various crystalline and polymorphic forms of Compound(I) may be used. As used herein, “polymorphic forms” or “polymorphs” ofcompound (I) is intended to encompass crystalline hydrates or othercrystalline solvates of compound (I).

Any chemical formula given herein is also intended to representunlabeled forms as well as isotopically labeled forms of the compounds.Isotopically labeled compounds have structures depicted by the formulaegiven herein except that one or more atoms are replaced by an atomhaving a selected atomic mass or mass number. Isotopes that can beincorporated into compounds of the disclosure include, for example,isotopes of hydrogen, carbon, nitrogen, and oxygen, such as ³H, ¹¹C,¹³C, ¹⁴C, and ¹⁵N. Accordingly, it should be understood that methods ofthe present invention can or may involve compounds that incorporate oneor more of any of the aforementioned isotopes, including for example,radioactive isotopes, such as ³H and ¹⁴C, or those into whichnon-radioactive isotopes, such as ²H and ¹³C are present. Suchisotopically labelled compounds are useful in metabolic studies (with¹⁴C), reaction kinetic studies (with, for example ²H or ³H), detectionor imaging techniques, such as positron emission tomography (PET) orsingle-photon emission computed tomography (SPECT) including drug orsubstrate tissue distribution assays, or in radioactive treatment ofpatients. Isotopically-labeled compounds can generally be prepared byconventional techniques known to those skilled in the art, e.g., usingan appropriate isotopically-labeled reagents in place of the non-labeledreagent previously employed.

The present invention encompasses embodiments that include allpharmaceutically acceptable salts of the compounds useful according tothe invention provided herein. As used herein, “pharmaceuticallyacceptable salt” refers to derivatives of the disclosed compoundswherein the parent compound is modified by converting an existing acidor base moiety to its salt form. Examples of pharmaceutically acceptablesalts include, but are not limited to, mineral or organic acid salts ofbasic residues such as amines; alkali or organic salts of acidicresidues such as carboxylic acids; and the like. The pharmaceuticallyacceptable salts include the conventional non-toxic salts of the parentcompound formed, for example, from non-toxic inorganic or organic acids.The pharmaceutically acceptable salts can be synthesized from the parentcompound which contains a basic or acidic moiety by conventionalchemical methods. Generally, such salts can be prepared by reacting thefree acid or base forms of these compounds with a stoichiometric amountof the appropriate base or acid in water or in an organic solvent, or ina mixture of the two; generally, non-aqueous media like ether, ethylacetate, ethanol, isopropanol, or acetonitrile are preferred. Lists ofsuitable salts are found in Remington's Pharmaceutical Sciences, 17^(th)ed., Mack Publishing Company, Easton, Pa., 1985, p. 1418 and Journal ofPharmaceutical Science, 66, 2 (1977), each of which is incorporatedherein by reference in its entirety. For example, preferredpharmaceutically acceptable salts include, but are not limited to,mineral or organic acid salts of basic residues such as amines. Forexample, the salt can be a hydrochloride salt. Other examples ofsuitable salts can be found in U.S. Pat. No. 8,349,852, the content ofwhich is hereby incorporated by its entirety.

The phrase “pharmaceutically acceptable” as employed herein refers tothose compounds, materials, compositions, and/or dosage forms which are,within the scope of sound medical judgment, suitable for use in contactwith the tissues of human beings and animals without excessive toxicity,irritation, allergic response, or other problem or complication,commensurate with a reasonable benefit/risk ratio.

Unless indicated otherwise, all ingredient concentrations are presentedin units of % weight/volume (% w/v). As is commonly understood, the %w/v value refers to the amount of the particular component or ingredientin the formulation. It is commonly understood that equivalentconcentrations can be expressed in different units. For example, aconcentration of 0.1% w/v can also be expressed as a 1 mg/ml solution.

Unless otherwise specified, the weight or dosage referred to herein forthe compound of formula I is the weight or dosage of the compounditself, not that of a salt or prodrug thereof, which can be different toachieve the intended therapeutic effect. For example, the weight ordosage of a corresponding salt of a compound suitable for the methods,compositions, or combinations disclosed herein may be calculated basedon the ratio of the molecular weights of the salt and compound itself.

The present invention provides formulations of the compound of formulaI. In some embodiments, the formulations are aqueous suspensions of acompound of formula I. In some embodiments, the suspension includes thecompound of formula I at a concentration of from about 0.5% to about1.5% w/v, about 0.5% to about 2.5% w/v, about 0.5% to about 3.5% w/v,about 0.5% to about 3.0% w/v, about 1.0% to about 2.5% w/v, about 1.5%to about 3.0% w/v, about 0.5% to about 2.5% w/v. In some embodiments,the concentration of the compound of formula I in a formulation fortopical ocular use is at least about 0.5% w/v, at least about 1.0% w/v,at least about 1.5% w/v, at least about 2.0% w/v, or at least about 2.5%w/v. In some embodiments, the concentration of the compound of formula Iin a formulation for topical use is no more than about 5.0% w/v, no morethan about 4.5% w/v, no more than about 4.0% w/v, no more than about3.5% w/v, or no more than about 3.0% w/v. In particular embodiments, theconcentration of the compound of formula I in a formulation for topicaluse is about 0.5% w/v, about 1.0% w/v, about 1.5% w/v, about 2.0% w/v,about 2.5% w/v, about 3.0% w/v, or about 3.5% w/v. Expressed in units ofmg/ml, in some embodiments, compound of formula I is administered to thesubject at a concentration of about 5 mg/ml to about 35 mg/ml, about 5mg/ml to about 25 mg/ml, or about 5 mg/ml to about 15 mg/ml, about 5mg/ml to about 30 mg/ml, about 10 mg/ml to about 25 mg/ml, about 15mg/ml to about 30 mg/ml, or about 5 mg/ml to about 25 mg/ml. In someembodiments, the concentration of the compound of formula I in aformulation for topical use is at least about 5 mg/ml, at least about 10mg/ml, at least about 15 mg/ml, at least about 20 mg/ml, or at leastabout 25 mg/ml. In some embodiments, the concentration of the compoundof formula I in a formulation for topical use is no more than about 50mg/ml, no more than about 45 mg/ml, no more than about 40 mg/ml, no morethan about 35 mg/ml, or no more than about 30 mg/ml. In particularembodiments, the compound of formula I is administered at aconcentration of about 5 mg/ml, about 10 mg/ml, about 15 mg/ml, about 20mg/ml, about 25 mg/ml, about 30 mg/ml, or about 35 mg/ml.

In some embodiments, the compound of formula is present as polymorphform B, as described in U.S. Pat. No. 8,349,852, incorporated byreference herein. In some embodiments, polymorph B is characterized byan X-ray diffraction pattern having three or more peaks at 2θ valuesselected from 9.3, 10.6 and 14.4±0.2° 2θ, when recorded using CuK_(α)radiation. In some embodiments, polymorph B is characterized by an X-raydiffraction pattern having three or more peaks at 2θ values selectedfrom 9.3, 10.6, 14.4, 15.5, 17.9, 19.9, 23.4±0.2° 2θ, when recordedusing CuK_(α) radiation. In some embodiments, polymorph B ischaracterized by an X-ray diffraction pattern having three or more peaksat 2θ values selected from 9.3, 10.6, 12.8, 14.4, 15.5, 17.9, 19.9,21.3, 23.4, and 28.0±0.2° 2θ, when recorded using CuK_(α) radiation.

In some embodiments of the formulations described herein, the D₉₀ ofcompound I in the formulation (diameter at which 90% of compound I iscomprised of smaller particles) is below about 10 μm, below about 8 μm,below about 6 μm, below about 4 μm, below about 3 μm, or about 2 μm. Insome embodiments, the D₅₀ of compound I in the formulation (diameter atwhich 50% of compound I is comprised of smaller particles) is belowabout 10 μm, below about 8 μm, below about 6 μm, below about 4 μm, belowabout 3 μm, below about 2 μm, or about 1 μm. In some embodiments, theD₁₀ of compound I in the formulation (diameter at which 10% of compoundI is comprised of smaller particles) is below about 5 μm, below about 4μm, below about 3 μm, below about 2 μm, below about 1 μm, or about 0.3μm.

In some embodiments, the formulation further includes at least oneophthalmically acceptable excipient.

In some embodiments, the formulations include an ophthalmicallyacceptable surfactant. In some embodiments, the surfactant is an anionicsurfactant. In specific embodiments, the anionic surfactant is selectedfrom C₁₀-C₂₂ alkylsulfates, C₁₀-C₂₂alkyl(oligooxyalkylene)sulfates,C₄-C₂₂ alkyl sulfosuccinate esters, C₁₀-C₂₂acylsarcosinates and C₁₀-C₂₂alkylcarboxylates; wherein oligooxyalkylene moieties have from one tofive oxy-C₁-C₆ alkylene moieties, e.g., oxyethylene moieties. Theanionic surfactants may have a countercation is selected from alkalimetal, e.g., sodium, C₁-C₃ alkylammonium, tri(C₁-C₃ alkanol)ammonium,e.g., triethanolammonium, C₃ alkanol)ammonium and ammonium cations. Theconcentration of anionic surfactant in the formulation is from about0.005 to 0.1 g/L, or 0.005 to 0.05 g/L. In some embodiments, thesurfactant is a cationic surfactant. Non-limiting examples of cationicsurfactants include alkylamine salts, alkylamine polyoxyethylene adduct,a fatty acid triethanolamine monoester salt, acyl aminoethyldiethylamine salts, fatty acid polyamine condensates, alkylimidazolines, 1-acyl aminoethyl-2-alkyl imidazoline,1-hydroxyethyl-2-alkyl imidazoline, include chlorhexidine or the likesalts thereof, chlorhexidine or a salt thereof, e.g., chlorhexidinegluconate. In some embodiments, the cationic surfactant is present inthe formulation in an amount of from about 0.001 to about 5% w/v, orabout 0.001 to about 1% w/v, or about 0.001 to about 0.1% w/v, or about0.001 to about 0.01% w/v, or about 0.001 to about 0.005% w/v.

In specific embodiments, the surfactant is a non-ionic surfactant. Insome embodiments, the non-ionic surfactant is a polysorbate surfactant,a block copolymer of ethylene oxide and propylene oxide surfactant(e.g., a pluronic or tetronic surfactant), poloxamer, tyloxapol, orcombinations thereof. Tyloxapol is a nonionic liquid polymer of thealkyl aryl polyether alcohol type. Poloxamers are nonionic triblockcopolymers composed of a central hydrophobic chain of polyoxypropylene(poly(propylene oxide)) flanked by two hydrophilic chains ofpolyoxyethylene (poly(ethylene oxide)). In particular embodiments, thenon-ionic surfactant is tyloxapol. In some embodiments, the tyloxapol ispresent in an amount at least about 0.001% w/v, at least about 0.01%w/v, at least about 0.02% w/v, at least about 0.03% w/v, at least about0.04% w/v, and no more than about 1% w/v, no more than about 0.5% w/v,no more than about 0.3% w/v, or no more than about 0.2% w/v, no morethan about 0.1% w/v, or no more than about 0.08% w/v. In particularembodiments, the non-ionic surfactant is tyloxapol, present in an amountof about 0.03% w/v to 0.08% w/v, or about 0.05% w/v. In particularembodiments, the surfactant is substantially all tyloxapol.

In some embodiments, the formulations include about 15-20% w/v of apoloxamer surfactant. In some embodiments, the formulations includeabout 15, about 15.5%, about 16%, about 16.5%, about 17%, about 17.5%,about 18%, about 18.5%, about 19%, about 19.5%, or about 20% w/v ofpoloxamer. In particular embodiments, the formulations include about 17%to about 18% w/v, or about 17.5% w/v poloxamer. In yet particularembodiments, the poloxamer is Poloxamer 407. In particular embodiments,the surfactant is substantially all tyloxapol.

In some embodiments, the formulations include a suspending agent. Insome embodiments, the suspending agent is a carbomer, hydroxypropylmethyl cellulose (hypromellose), polyethylene glycol, or combinationsthereof. Carbomers are carboxyvinyl polymers that have a network ofcross-linked polymer chains. The polymers are often characterized ashaving carboxylic acid functional groups and preferably contain from 2to 7 carbon atoms per functional group. Carbomers, i.e. synthetichigh-molecular-weight polymers of acrylic acid that are crosslinked e.g.with allyl sucrose or allyl ethers of pentaerythritol, particularlywater-soluble and water-swellable carbomers. Carbomers are availableunder the trade name CARBOPOL® from various suppliers In particularembodiments, the carbomer is carbomer homopolymer Type B. In particularembodiments, the carbomer is CARBOPOL® 934P (Carbomer 934P), 940 or974P. In some embodiments, the suspending agent is carbomer and ispresent in the formulation in an amount of at least about 0.05% w/v, atleast about 0.1% w/v, or at least about 0.2% w/v, and no greater thanabout 1.0% w/v, no greater than about 0.6% w/v, or no greater than about0.5%. In particular embodiments, the suspending agent is carbomer, andis present in the formulation in an amount of 0.1% w/v to about 0.3%w/v, or about 0.2% w/v.

In some embodiments, the suspending agent is hydroxypropyl methylcellulose. In particular embodiments, the hydroxypropyl methyl celluloseis present in the formulation in an amount of at least about 0.05% w/v,at least about 0.1% w/v, or at least about 0.25% w/v, and less thanabout 1.8% w/v, less than about 1.0% w/v, less than about 0.8% w/v, orless than about 0.6% w/v. In some embodiments, the hydroxypropyl methylcellulose is present in the formulation in an amount of from about 0.1%w/v to about 0.8 w/v %; from about 0.1% w/v to about 0.6% w/v; fromabout 0.25% w/v to about 0.8% w/v; from about 0.4% w/v to about 0.6%w/v.

In some embodiments, the suspending agent is a polyethylene glycol (PEG)having molecular weight of at least about 200 Da. In some embodiments,the PEG has a molecular weight of at least about 400, 1,000, 2,000,3,000, 4,000, 6,000, or about 10,000 Da. In some embodiments, thesuspending agent is a polyethylene glycol (PEG) having molecular weightof from about 200 to about 20,000 Da. In some embodiments, the PEG has amolecular weight of about 400, 1,000, 2,000, 3,000, 4,000, 6,000, orabout 10,000 Da. In some embodiments, the PEG is present in theformulation in an amount of at least 1% w/v, at least about 2% w/v, atleast about 3% w/v, and less than about 10% w/v, less than about 9% w/v,or less than about 8% w/v. In particular embodiments, the suspendingagent is PEG400 at a concentration of from about 4% w/v to about 9% w/v,about 5% w/v to about 8% w/v, or about 7% w/v. In particularembodiments, the suspending agent is PEG6000 at a concentration of fromabout 1% w/v to about 4% w/v, about 1% w/v to about 3% w/v, or about 2%w/v.

In particular embodiments, the suspending agent is a combination of morethan one suspending agent. In other embodiments, the suspending agent issubstantially all carbomer.

In some embodiments, the suspending agent can provide the desiredviscosity of the formulation. Without being bound by theory, it isbelieved that suspending agents may increase the viscosity of theformulation. A formulation with appropriate viscosity is beneficial inmaintaining compound I in a suspended state in the formulation withoutsettling and caking. In some embodiments, the formulation viscosity isfrom about 10 to about 200 cP (centipoise), from about 20 cP to about200 cP, or from about 20 cP to about 150 cP. In some embodiments, theformulation viscosity is at least about 10 cP, 20 cP, 50 cP, 100 cP, orat least about 150 cP. Viscosity measurements for the formulations aremeasured using a Brookfield viscometer using spindle CP-42 at either 3rpm or 60 rpm. Viscosity is typically measured at room temperature,i.e., 25° C.

In some embodiments, the formulation includes a tonicity agent. In someembodiments, the tonicity agent is a polyol. As used herein, the term“polyol” includes any compound having at least one hydroxyl group oneach of two adjacent carbon atoms that are not in trans configurationrelative to each other. The polyols can be linear or cyclic, substitutedor unsubstituted, or mixtures thereof, so long as the resultant complexis water soluble and pharmaceutically acceptable. Examples of suchcompounds include: sugars, sugar alcohols, sugar acids and uronic acids.In some embodiments, the tonicity agent is a polyol such as sugars,sugar alcohols and sugar acids, including, but not limited to: mannitol,glycerin, xylitol, sorbitol and propylene glycol, or combinationsthereof. In particular embodiments, the composition includes mannitol,glycerin or a combination thereof. In some embodiments, the amount ofpolyol in the formulation is from about 0.05% w/v to about 10% w/v, fromabout 0.1% to about 8% w/v, from about 0.1% to about 7% w/v, from about0.1% to about 5% w/v. In particular embodiments, the tonicity agent ismannitol or glycerin, which is present in the formulation in an amountof from 0.1% w/v to about 5% w/v, or about 0.2% w/v, about 0.3% w/v,about 0.4% w/v, about 0.5% w/v, about 1% w/v, about 2% w/v, about 2.5%w/v, about 3.0% w/v, about 3.5% w/v, about 4.0% w/v, about 4.5% w/v, orabout 5% w/v. In particular embodiments, the tonicity agent is mannitol.In particular embodiments, the tonicity agent is glycerin.

In some embodiments, the formulation includes a buffer. Non limitingexamples of buffer substances include acetate, ascorbate, borate,hydrogen carbonate, carbonate, citrate, edetate (EDTA) gluconate,lactate, phosphate, propionate and TRIS (tromethamine) buffers. Inparticular embodiments, the buffer is a phosphate buffering system. Inparticular embodiments, the buffer is a tromethamine buffer. The amountof buffer substance added is, typically, that necessary to ensure andmaintain a physiologically tolerable pH range. In some embodiments, thepH range is in the range of from about 4 to about 9, from about 4.5 toabout 8.5, from about 5.0 to about 8.0, from about 5.5 to about 8.0,from about 6.4 to about 8.4. In some embodiments, the pH is about 6.0.In particular embodiments, the pH is about 7.4. In some embodiments, thepH of the formulation is about 5.0 to about 8.0, about 5.5 to about 7.5,about 5.0 to about 7.4, about 5.5 to about 7.4, about 6.0 to about 8.0,about 6.5 to about 8.0, about 6.0 to about 7.4, or about 6.5 to about7.4.

In some embodiments, the formulation includes a salt. In someembodiments, salt is sodium chloride, potassium chloride, calciumchloride, or magnesium chloride. In particular embodiments, the salt issodium chloride. In particular embodiments, the salt is present in anamount of at least about 0.01% w/v, at least about 0.02% w/v, at leastabout 0.03% w/v, at least about 0.04% w/v, and no more than about 0.5%w/v, no more than about 0.4% w/v, no more than about 0.3% w/v, no morethan about 0.2% w/v, or no more than about 0.1% w/v. In particularembodiments, the salt is present in an amount of from about 0.01% w/v toabout 0.5% w/v, from about 0.02% w/v to about 0.4% w/v, from about 0.03%w/v to about 0.3% w/v, from about 0.04% w/v to about 0.2% w/v, fromabout 0.05% w/v to about 0.1% w/v. In particular embodiments, the saltis sodium chloride and is present in the formulation in an amount ofabout 0.02% to about 0.07% w/v, or about 0.05% w/v.

In some embodiments, the formulations described herein have anosmolality of about 200 to about 450 milliosmoles per kilogram(mOsm/kg), about 200 to about 400 mOsm/kg, about 200 to about 300mOsm/kg, or about 240 to about 360 mOsm/kg.

In some embodiments, the formulation may also be self-preserved and doesnot include a preservative. In other embodiments, the formulationincludes a preservative. In some embodiments, the preservative includes,without limitation, polyhexylmethylene biguanidine (PHMB), polymericquaternary ammonium compound (e.g., polyquaternium-1), chlorinecontaining preservatives such as benzalkonium chloride (BAK), chloritepreservatives or others.

In some embodiments, the preservative is polymeric quaternary ammoniumcompounds that are ophthalmically acceptable. Compounds of this type aredescribed in U.S. Pat. Nos. 3,931,319; 4,027,020; 4,407,791; 4,525,346;4,836,986; 5,037,647 and 5,300,287; and PCT application WO 91/09523(Dziabo et al.). In particular embodiments, the polymeric ammoniumcompound is polyquaternium 1, otherwise known as POLYQUAD® or ONAMERM®with a number average molecular weight between 2,000 to 30,000. In stillparticular embodiments, the number average molecular weight is between3,000 to 14,000.

When used, the polymeric quaternary ammonium compound is generally usedin an amount that is greater than about 0.00001 w/v %, greater thanabout 0.0003 w/v %, or greater than about 0.0007 w/v % of theformulation. Moreover, the polymeric quaternary ammonium compound, whenused in the formulation, is generally used at a concentration that isless than about 0.03 w/v %, less than about 0.003 w/v %, or less thanabout 0.0015 w/v % of the formulation. In some embodiments, theconcentration of polymeric quaternary ammonium compound in theformulation are as follows: greater than about 0.0003 w/v % but lessthan about 0.003 w/v %; greater than about 0.0003 w/v % but less thanabout 0.0015 w/v %; greater than about 0.0007 w/v % but less than about0.003 w/v %; and greater than about 0.0007 w/v % but less than about0.0015 w/v %. In particular embodiments, the formulation includespolyquarternium 1 at a concentration of about 0.001% w/v.

In some embodiments, the formulation includes BAK at a concentrationthat is at least about 0.0005 w/v %, about 0.001 w/v %, or greater thanabout 0.007 w/v % of the formulation, and at a concentration that isless than about 0.1 w/v %, less than about 0.02 w/v %, or less thanabout 0.0035 w/v % of the ophthalmic composition. It is specificallycontemplated that any of the lower limits on the concentration of BAKmay be used in conjunction with any of the upper limits on theconcentrations of BAK. In particular embodiments, the concentration ofBAK in the composition are as follows: greater than about 0.001 w/v %but less than about 0.02 w/v %; greater than about 0.001 w/v % but lessthan about 0.0035 w/v %; greater than about 0.007 w/v % but less thanabout 0.02 w/v %; and greater than about 0.007 w/v % but less than about0.0035 w/v %.

In some embodiments, described herein is an aqueous formulation thatincludes:

4-(7-hydroxy-2-isopropyl-4-oxo-4H-quinazolin-3-yl)-benzonitrile(compound I) or a salt, co-crystal, or polymorph thereof, in an amountof about 0.5% w/v to about 3.5% w/v,

and one or more excipients selected from the group consisting of asurfactant, a suspending agent, a tonicity agent, a buffer, apreservative, a salt, and a preservative.

In some embodiments, described herein is an aqueous formulation thatincludes:

4-(7-hydroxy-2-isopropyl-4-oxo-4H-quinazolin-3-yl)-benzonitrile(compound I) or a salt, co-crystal, or polymorph thereof, present as asuspension in the formulation, in an amount of about 0.5% w/v to about3.5% w/v,

and one or more excipients selected from the group consisting of asurfactant, a suspending agent, a tonicity agent, a buffer, apreservative, a salt, and a preservative.

In some embodiments, described herein is an aqueous formulation thatincludes:

4-(7-hydroxy-2-isopropyl-4-oxo-4H-quinazolin-3-yl)-benzonitrile(compound I) or a salt, co-crystal, or polymorph thereof, present as asuspension in the formulation,

and one or more excipients selected from the group consisting of asurfactant, a suspending agent, a tonicity agent, a buffer, apreservative, a salt, and a preservative.

In some embodiments, described herein is an aqueous formulation thatincludes:

a suspension of4-(7-hydroxy-2-isopropyl-4-oxo-4H-quinazolin-3-yl)-benzonitrile(compound I) or a salt, co-crystal, or polymorph thereof,

and one or more excipients selected from the group consisting of asurfactant, a suspending agent, a tonicity agent, a buffer, apreservative, a salt, and a preservative.

In some embodiments, described herein is an aqueous formulation thatincludes:

4-(7-hydroxy-2-isopropyl-4-oxo-4H-quinazolin-3-yl)-benzonitrile(compound I) or a salt, co-crystal, or polymorph thereof, in an amountof about 0.5% w/v to about 3.5% w/v, present as a suspension in theformulation

a surfactant,

a suspending agent,

and one or more excipients selected from the group consisting of atonicity agent, a buffer, a preservative, a salt, and a preservative.

In some embodiments, the invention described herein is a formulationthat includes:

4-(7-hydroxy-2-isopropyl-4-oxo-4H-quinazolin-3-yl)-benzonitrile(compound I) or a salt, co-crystal, or polymorph thereof, in an amountof about 0.5% w/v to about 3.5% w/v, present as a suspension in theformulation,

a non-ionic surfactant;

a suspending agent;

a tonicity agent;

a buffer;

a salt; and

optionally, a preservative.

In some embodiments, the invention described herein is a formulationthat includes:

a suspension of4-(7-Hydroxy-2-isopropyl-4-oxo-4H-quinazolin-3-yl)-benzonitrile(compound I) or a salt, co-crystal, or polymorph thereof, in an amountof about 0.5% w/v to about 3.5% w/v,

a non-ionic surfactant;

a suspending agent;

a tonicity agent;

a buffer;

a salt;

optionally, a preservative; and

water qs to 100%.

In some embodiments, the invention described herein is a formulationthat includes:

a suspension of4-(7-Hydroxy-2-isopropyl-4-oxo-4H-quinazolin-3-yl)-benzonitrile(compound I) or a salt, co-crystal, or polymorph thereof, in an amountof about 0.5% w/v to about 2.5% w/v,

a non-ionic surfactant selected from tyloxapol, poloxamer, orcombinations thereof in an amount of from about 0.01 to 0.2% w/v;

a suspending agent selected from hydroxypropyl methyl cellulose,polyethylene glycol or carbomer homopolymer Type B;

a tonicity agent selected from polyols in an amount of from about 0.05%w/v to about 10% w/v;

a buffer selected from edetate, phosphate, borate, or combinationsthereof;

a salt; and

water qs to 100%; and

a pH in the range of from about 5.5 to about 8.0.

In some embodiments, the invention described herein is a formulationthat includes:

a suspension of4-(7-Hydroxy-2-isopropyl-4-oxo-4H-quinazolin-3-yl)-benzonitrile(compound I) or a salt, co-crystal, or polymorph thereof, in an amountof about 0.5% w/v, about 1.0% w/v, about 1.5% w/v, about 2.0% w/v, orabout 2.5% w/v,

a non-ionic surfactant selected from tyloxapol in an amount of about0.04 w/v to about 0.06% w/v, poloxamer in an amount of about 0.005-0.12%w/v, or combinations thereof;

a suspending agent selected from hydroxypropyl methyl cellulose in anamount of from about 0.1% w/v to about 0.8 w/v %, polyethylene glycol inan amount of from about 2% w/v to about 8% w/v, carbomer homopolymerType B in an amount from about 0.05% w/v to about 0.5% w/v, orcombinations thereof;

a tonicity agent selected from mannitol or glycerin in an amount of fromabout 0.1% w/v to about 5% w/v;

a buffer selected from edetate, phosphate, borate, tromethamine, orcombinations thereof;

sodium chloride in an amount of from 0.01% w/v to about 1% w/v;

water qs to 100% and

a pH in the range of from about 5.5 to about 8.0.

In some embodiments, the invention described herein is a formulationthat includes:

a suspension of4-(7-Hydroxy-2-isopropyl-4-oxo-4H-quinazolin-3-yl)-benzonitrile(compound I) or a salt, co-crystal, or polymorph thereof, in an amountof about 0.5% w/v, about 1.0% w/v, about 1.5% w/v, about 2.0% w/v, orabout 2.5% w/v,

tyloxapol in an amount of about 0.04 w/v to about 0.06% w/v;

carbomer homopolymer Type B in an amount from about 0.05% w/v to about0.4% w/v;

glycerin in an amount of from about 0.5% w/v to about 5% w/v;

a buffer selected from edetate, phosphate, borate, tomethamine, orcombinations thereof;

sodium chloride in an amount of from 0.01% w/v to about 1% w/v;

water qs to 100% and

a pH in the range of from about 5.5 to about 8.0.

In some embodiments, the invention described herein is a formulationthat includes:

a suspension of polymorphic form B of4-(7-Hydroxy-2-isopropyl-4-oxo-4H-quinazolin-3-yl)-benzonitrile(compound I), in an amount of about 0.5% w/v, about 1.0% w/v, about 1.5%w/v, about 2.0% w/v, or about 2.5% w/v,

tyloxapol in an amount of about 0.04 w/v to about 0.06% w/v;

carbomer homopolymer Type B in an amount from about 0.05% w/v to about0.4% w/v;

glycerin in an amount of from about 0.5% w/v to about 5% w/v;

a buffer selected from edetate, phosphate, borate, tomethamine, orcombinations thereof;

sodium chloride in an amount of from 0.01% w/v to about 1% w/v;

water qs to 100% and

a pH in the range of from about 5.5 to about 8.0.

In some embodiments, the invention described herein is a formulationthat includes:

a suspension of polymorphic form B of4-(7-Hydroxy-2-isopropyl-4-oxo-4H-quinazolin-3-yl)-benzonitrile(compound I), in an amount of about 0.5% w/v, about 1.0% w/v, about 1.5%w/v, about 2.0% w/v, or about 2.5% w/v,

about 0.05% w/v of tyloxapol;

about 0.2% w/v of carbomer homopolymer Type B;

about 2.0% of glycerin;

a tromethamine buffer;

about 0.05% w/v of sodium chloride; and

water qs to 100% and

a pH in the range of from about 6.4 to about 8.4;

wherein the formulation does not include a preservative.

In some embodiments, acids or bases such as hydrochloric acid, sodiumhydroxide, or combinations thereof are used to adjust pH of theformulation. In particular embodiments, hydrochloric acid is used toadjust pH to about 6.0, or about 7.4.

In some embodiments, the formulations described herein are aqueous, thatis, they include at least about 90%, at least about 92%, or at leastabout 95% water.

Without being bound by theory, it is believed that the viscosity offormulations with carbomer homopolymer Type B increases as the pH isadjusted from acidic to neutral, but the viscosity decreases withincreasing ionic strength. The present inventors found that usingtromethamine as the buffer increases pH without substantial increases inionic strength. Without further being bound by theory, the inventorsalso found that glycerin is able adjust the tonicity of the formulationwithout increasing its ionic strength. Glycerin is also well-toleratedand non-irritating and also serves as a humectant and an additionalviscosity agent.

Without being bound by any particular theory, the present inventors alsofound that inclusion of a surfactant in the formulation acts as awetting agent for the compound of formula I, thereby providing adequatewetting of the particles of compound I while decreasing the potentialfor irritation and foaming and aiding in redispersibility of thesuspension.

In some embodiments, the formulations described herein further includeadditional components. In particular embodiments, the formulationincludes a cyclodextrin derivative, for example, β-cyclodextrinderivative, γ-cyclodextrin derivative or a combination thereof. Inparticular embodiments, the cyclodextrin is a hydroxypropylβ-cyclodextrin or sulfoalkylether β-cyclodextrin. When present, thecyclodextrin derivative may be present in an amount of at least about1.5 w/v %, at least about 3.0 w/v %, at least about 3.5 w/v % or atleast about 4.5 w/v, but no greater than about 10.0 w/v %, no greaterthan about 8.0% w/v, no greater than about 6.5 w/v %, or no greater thanabout 5.5 w/v. In particular embodiments, the formulation includes about5% w/v of either hydroxypropyl β-cyclodextrin or sulfoalkyletherβ-cyclodextrin.

In some embodiments, the formulations of the invention may include anadditional therapeutic agent in addition to compound (I). Furthertherapeutic agents may include, for instance, other compounds andantibodies useful for treating ocular surface disorders. A non-limitinglist of such agents incudes nonsteroidal anti-inflammatory drugs such asketorolac, nepafenac, bromfenac, corticosteroids; drugs for dry eyedisease such as cyclosporine, lifitegrast, or other TRPV1 inhibitors. Inparticular embodiments, the additional therapeutic agent is anophthalmic steroid such as dexamethasone, fluocinolone, loteprednol,difluprednate, fluorometholone, prednisolone, prednisone, medrysone,triamcinolone, betamethasone, rimexolone, or pharmaceutically acceptablesalts thereof. Further non-limiting examples of such additionaltherapeutic agents that may be included in the pharmaceuticalcomposition include Xiidra® (lifitegrast), Restasis® (cyclosporine),minocycline, doxycycline, or other tetracycline antibiotics. Otherexamples include keratolytic agents such as selenium disulfide,salicylic acid, glycolic acid etc., or pharmaceutically acceptable saltsthereof.

In some embodiments, the formulation is stored at refrigeratedtemperatures (e.g., 4° C.). In some embodiments, the formulation iswarmed to room temperature prior to administration.

In some embodiments, the suspension is packaged in a single dosecontainer. In some embodiments, the formulation is packaged in amulti-dose container.

The formulations described herein are delivered to the surface of theeye one to six times a day, depending on the routine discretion of theskilled clinician. In some embodiments, the formulations areadministered, one, two, three, or four times a day.

In some embodiments, the formulation exhibits settling of less thanabout 10% after storage at room temperature for six months. In someembodiments, the formulation exhibits settling of less than about 8%,less than about 7%, less than about 6%, less than about 5%, less thanabout 4%, less than about 3%, or less than about 2% after storage atroom temperature for six months. Settling behavior is measured bymethods commonly known to those of skill in the art, for example asdescribed herein.

In some embodiments, the amount of compound I in the formulation is atleast 90% of the initial amount after about 6 months of storage underrefrigeration (e.g., about 4° C.). In some embodiments, the amount ofcompound I in the formulation is at least about 90% of the initialamount after about 8 months, about 10 months, about 12 months, about 15months, or about 18 months of storage under refrigeration. In someembodiments, the amount of compound I in the formulation is at leastabout 91%, at least about 92%, at least about 93%, at least about 94%,at least about 95%, at least 96%, at least about 97% or at least about98% of the initial amount after about 6 months of storage underrefrigeration (e.g., about 4° C.). In some embodiments, the amount ofcompound I in the formulation at least about 91%, at least about 92%, atleast about 93%, at least about 94%, at least about 95%, at least 96%,at least about 97% or at least about 98% of the initial amount afterabout 18 months of storage under refrigeration. The amount of compound Iin the formulation is measured using methods commonly known to those ofskill in the art, for example, HPLC, LC/MS, etc.

In some embodiments, the formulation comprises no more than about 10% ofa degradation product after 6 months under refrigeration, wherein thedegradation product has a relative retention time of 1.23, compared tocompound I, when analyzed by HPMC using a gradient 0.1% trifluoroaceticacid (TFA) water/acetonitrile mobile phase. In some embodiments, theformulation comprises no more than about 9%, no more than about 8%, nomore than about 7%, no more than about 6%, no more than about 5%, nomore than about 4%, no more than about 3%, or no more than about 2% of adegradation product after 6 months under refrigeration.

In some embodiments, no more than about 10% of compound I in theformulation degrades upon storage for 12 weeks at 40° C. In particularembodiments, no more than about 9%, no more than about 8%, no more thanabout 7%, no more than about 6%, no more than about 6%, no more thanabout 5%, no more than about 4%, no more than about 3%, no more thanabout 2% of compound I in the formulation degrades upon storage for 12weeks at 40° C.

In some embodiments, the pharmaceutical formulations of the inventionmay include an additional therapeutic agent in addition to Compound (I).Further therapeutic agents may include, for instance, other compoundsand antibodies useful for treating ocular surface disorders. Anon-limiting list of such agents incudes nonsteroidal anti-inflammatorydrugs such as ketorolac, nepafenac, bromfenac, corticosteroids; drugsfor dry eye disease such as cyclosporine, lifitegrast, autologous serum,or other TRPV1 inhibitors. In particular embodiments, the additionaltherapeutic agent is an ophthalmic steroid such as dexamethasone,fluocinolone, loteprednol, difluprednate, fluorometholone, prednisolone,prednisone, medrysone, triamcinolone, betamethasone, rimexolone, orpharmaceutically acceptable salts thereof. Further non-limiting examplesof such additional therapeutic agents that may be included in thepharmaceutical composition include Xiidra® (lifitegrast), Restasis®(cyclosporine), minocycline, doxycycline, or other tetracyclineantibiotics. Other examples include keratolytic agents such as seleniumdisulfide, salicylic acid, glycolic acid etc., or pharmaceuticallyacceptable salts thereof.

Methods of Making

In some embodiments, the present invention is a method of making thepharmaceutical formulations of compound I.

In some embodiments, the formulation is prepared by mixing an amount of4-(7-hydroxy-2-isopropyl-4-oxo-4H-quinazolin-3-yl)-benzonitrile(compound I) or a salt, co-crystal, or polymorph thereof, and one ormore excipients selected from the group consisting of a surfactant, asuspending agent, a tonicity agent, a buffer, a preservative, a salt,and a preservative.

In some embodiments, the formulation is prepared by mixing an amount of4-(7-Hydroxy-2-isopropyl-4-oxo-4H-quinazolin-3-yl)-benzonitrile(compound I) or a salt, co-crystal, or polymorph thereof,

a non-ionic surfactant;

a suspending agent;

a tonicity agent;

a buffer;

a salt;

optionally, a preservative; and

water qs to 100%, and

adjusting the pH to a range of from about 5.5 to about 8.0.

In some embodiments, the compound I is added as a stock suspension inwater, optionally with the surfactant. In some embodiments, the compoundI is present in an amount of 10% in the stock suspension. In alternativeor additional embodiments, the stock suspension of compound I furtherincludes 0.2% tyloxapol. In some embodiments, the stock suspension ofcompound I is milled to achieve a desired particle size of compound I.In some embodiments, the D₉₀ of compound I in the formulation (diameterat which 90% of compound I is comprised of smaller particles) is belowabout 10 μm, below about 8 μm, below about 6 μm, below about 4 μm, belowabout 3 μm, or about 2 μm. In some embodiments, the D₅₀ of compound I inthe formulation (diameter at which 50% of compound I is comprised ofsmaller particles) is below about 10 μm, below about 8 μm, below about 6μm, below about 4 μm, below about 3 μm, below about 2 μm, or about 1 μm.In some embodiments, the D₁₀ of compound I in the formulation (diameterat which 10% of compound I is comprised of smaller particles) is belowabout 5 μm, below about 4 μm, below about 3 μm, below about 2 μm, belowabout 1 μm, or about 0.3 μm.

In some embodiments, a 10% w/v slurry of compound I is jar milled on aroller mill at about 60 rpm for about 2 hours or about 3 hours.

In particular embodiments, the non-ionic surfactant, the suspendingagent, the tonicity agent, buffer, and salt are as noted supra. In someembodiments, the formulation does not include a preservative.

An exemplary method of manufacture of a 1.5% w/v compound I suspensionis described below:

-   -   1. Tare a clean, dry glass Schott bottle with a polybutylene        terephthalate, PTFE-lined cap and a magnetic stir bar.    -   2. Add the batch quantity of compound I vehicle (including        suspending agent, tonicity agent, surfactant, and salt, and pH        adjusted to final pH) to the bottle. Seal and steam sterilize        the compounding vessel (F₀≥30).    -   3. Transfer the vessel to a horizontal laminar flow workbench        and allow to cool.    -   4. Aseptically weigh and add the batch quantity of sterile 10%        compound I/0.2% surfactant to the compounding vessel. Adjust to        final batch weight, if necessary, with sterile (either steam        sterilized or aseptically filtered) Purified Water qs and stir        until uniform.    -   5. Aseptically fill the 1.5% suspension into sterile dispensing        bottles. Insert suspension tips and tighten screw-on closures to        seat tips and seal.    -   6. Remove filled units from laminar flow workbench and label.        Measure the final pH and osmolality values.

Methods of Use

Without being bound by theory, it is hypothesized that blockers of theTransient Receptor Potential Vanilloid 1 (TRPV1) receptor may be usefulin the treatment of pain, e.g., chronic pain.

Accordingly, in some embodiments, the invention provides a method oftreating ocular surface pain in a subject in need thereof, said methodincludes administering to the subject an effective amount of compound(I), or a pharmaceutically acceptable salt, solvate, or co-crystalthereof. In some embodiments, the invention provides a method ofreducing ocular surface pain in a subject in need thereof, said methodincludes administering to the subject an effective amount of compound(I), or a pharmaceutically acceptable salt, solvate, or co-crystalthereof. In some embodiments, the invention provides for the use of thecompound of formula I, or a pharmaceutically acceptable salt, solvate,or co-crystal thereof, in the treatment of ocular surface pain. In someembodiments, the compound of formula I is in polymorphic form B. Inparticular embodiments, the methods described herein are carried out byadministering the formulations of compound I described supra. Thus, theinvention provides a method of treating ocular surface pain byadministering a formulation of compound I as described herein. Inparticular embodiments, the method is the administration of aformulation that includes compound I as polymorph B.

In some embodiments, the subject suffers from episodic or acute ocularpain. In some embodiments, the subject suffers from chronic ocularsurface pain, which lasts for at least three months. In someembodiments, the subject suffers from chronic ocular surface pain, whichlasts for at least two months. In some embodiments, the subject suffersfrom chronic ocular surface pain, which lasts for at least one month. Insome embodiments, the subject suffers from chronic ocular surface pain,which lasts for at least four months. In some embodiments, the subjectsuffers from chronic ocular surface pain, which lasts for at least fivemonths. Thus, in some embodiments, the invention provides a method oftreating chronic ocular surface pain in a subject by administering tothe subject an effective amount of compound of formula I, or a salt,solvate, polymorph, or co-crystal thereof. In some embodiments, theinvention provides a method of reducing chronic ocular surface pain in asubject by administering to the subject an effective amount of compoundof formula I, or a salt, solvate, polymorph, or co-crystal thereof. Theinvention provides for the use of the compound of formula I, or apharmaceutically acceptable salt, solvate, polymorph, or co-crystalthereof, in the treatment of chronic ocular surface pain. In someembodiments, the compound of formula I is in a formulation as describedherein. In particular embodiments, the method is the administration of aformulation that includes compound I as polymorph B.

In some embodiments, the formulation is administered to the ocularsurface of the subject, e.g., any part of the cornea, conjunctiva, or tothe cul de sac of the eye.

In some embodiments, the invention provides for the administration ofthe compound of formula I to a subject in need thereof in aophthalmically compatible formulation at a concentration of about 0.5%w/v to about 3.5% w/v. In some embodiments, concentrations foradministration range from about 0.5% to about 3.5% w/v, about 0.5% toabout 2.5% w/v, about 0.5% to about 1.5% w/v, about 0.5% to about 3.0%w/v, about 1.0% to about 2.5% w/v, about 1.5% to about 3.0% w/v, about0.5% to about 2.5% w/v. In some embodiments, the concentration of thecompound I in an ophthalmically compatible formulation is at least about0.5% w/v, at least about 1.0% w/v, at least about 1.5% w/v, at leastabout 2.0% w/v, or at least about 2.5% w/v. In some embodiments, theconcentration of the compound of formula I in a formulation for topicaluse is no more than about 5.0% w/v, no more than about 4.5% w/v, no morethan about 4.0% w/v, no more than about 3.5% w/v, or no more than about3.0% w/v. In particular embodiments, the concentration of the compoundof formula I in a formulation for topical use is about 0.5% w/v, about1.0% w/v, about 1.5% w/v, about 2.0% w/v, about 2.5% w/v, about 3.0%w/v, or about 3.5% w/v. In some embodiments, the dose per administrationper eye is from about 0.15 to about 1.15 mg, or about 0.15 mg, 0.2 mg,about 0.25 mg, about 0.3 mg, about 0.35 mg, about 0.4 mg, about 0.45 mg,about 0.5 mg, about 0.55 mg, about 0.6 mg, about 0.65 mg, about 0.7 mg,about 0.75 mg, about 0.8 mg, about 0.85 mg, about 0.9 mg, about 0.95 mg,about 1.0 mg, about 1.05 mg, about 1.1 mg, or about 1.15 mg. In someembodiments, the dose per administration per eye is about 0.18 mg, about0.37 mg, about 0.55 mg, about 0.74 mg, or about 0.92 mg. In someembodiments, the total daily dose per eye is about 0.5 to about 3.5 mg,or about 0.5 mg, about 1.0 mg, about 1.5 mg, about 2.0 mg, about 2.5 mg,about 3.0 mg, or about 3.5 mg. In some embodiments, the compound offormula I is administered to the subject one to six times a day, e.g.,one, two, three, or four times a day. In some embodiments, the compoundof formula I is administered to the subject for a period of at leastabout one month, at least about two months, or at least about threemonths. In some embodiments, the compound of formula I is administeredto the subject for a period of at least about 12 weeks.

In some embodiments, the ocular surface pain or the chronic ocularsurface pain is associated with one or more of dry eye disease,Sjogren's Syndrome, conjunctivitis (including keratoconjuctivitis,vernal keratoconjunctivitis, allergic conjunctivitis),Map-Dot-Fingerprint Dystrophy, acanthamoeba, fibromyalgia, Meibomiangland dysfunction, thyroid eye disease, rosacea, ptosis, keratoconus,ocular pain syndrome, Steven-Johnson's syndrome, cornealepitheliopathies, corneal neuropathies (including LASIK induced cornealneuropathies), corneal dystrophies (including recurrent cornealdystrophies), epithelial basement membrane dystrophy, corneal erosionsor abrasions (including recurrent corneal erosions or abrasions), ocularsurface diseases, blepharitis, graft vs host disease, meibomitis,glaucoma, conjunctivochalasis, keratopathis (including herpetickeratopathy, filamentary keratopathy, band or bullous keratopathy,exposure keratopathy), keratitis (including herpes simplex viruskeratitis), iritis, episclentis, corneal surgery, multiple sclerosis,trichiasis, pterygium, neuralgia, xerophthalmia, or patients recoveringfrom neurotrophic keratitis.

In particular embodiments, the ocular surface pain or the chronic ocularsurface pain is associated with dry eye disease or Sjogren's Syndrome.In some embodiments of the methods described herein, the subject suffersfrom ocular pain persisting for at least three months afterphotorefractive keratectomy (PRK) surgery or laser-assisted in situkeratomileusis (LASIK) surgery.

In some embodiments, the subject suffers from conjunctivitis,subconjunctival hemorrhage, subconjunctival scarring, conjunctivalmembranes, conjunctival ulceration, superficial punctate epithelialerosions, epithelial defects, lid margin ulceration, lid marginkeratinization, symblepharon, ankyloblepharon, trichiasis, anteriorblepharitis, punctal auto-occlusion, meibomian gland disease, cornealopacification, dry eye, districhiasis, limbal stem cell failure, orcorneal vascularization.

In some embodiments, the administration of compound of formula I resultsin a reduction in the subject's ocular pain, compared to a placebo. Insome embodiments, the reduction in the subject's ocular pain is at leastabout 3 when measured on a VAS score, compared to a placebo. In someembodiments, the administration results in a reduction in the subject'socular pain of at least about 4, at least about 5, at least about 6, atleast about 7, at least about 8, at least about 9, or at least about 10,when measured on the VAS score, compared to a placebo. In someembodiments, the administration results in a reduction in the subject'spain of at least about 10%, at least about 15%, at least about 20%, orat least about 25%, compared to a placebo, about one hour, about 2hours, or about 2-4 hours after the administration. In some embodiments,the administration results in a reduction in the subject's pain, whenmeasured after 7 days of administration of the compound of formula I. Insome embodiments, the administration results in a reduction in thesubject's pain, when measured after 14 days of administration of thecompound of formula I.

In some embodiments, the administration of the compound of formula Iresults in a reduction in the subject's pain of at least about 2compared to a placebo, as measured by the VAS score, about half hourafter the administration.

In some embodiments, the reduction in pain score arises from thedifference in pain scores prior to and after administration of compoundI to the subject. In some embodiments, the reduction in pain score asmeasured by the VAS, arises from the difference in VAS scores prior toand after administration of compound I to the subject. In someembodiments, the reduction in VAS score occurs within about half hourafter administration of compound I to the subject. In some embodiments,the reduction in pain score occurs within about 1 hour, about 2 hours,about 3 hours, about 4 hours, about 5 hours, or about 6 hours afteradministration of compound I to the subject. In some embodiments, theadministration results in a reduction in the subject's pain, whenmeasured after 7 days of administration of the compound of formula I. Insome embodiments, the administration results in a reduction in thesubject's pain, when measured after 14 days of administration of thecompound of formula I.

In some embodiments, the administration of the compound of formula Iresults in an improved score on at least one question of the OPAS of atleast about 10%, at least about 20%, or at least about 30%.

In some embodiments, the administration of the compound of formula Iresults in an improved score on at least one question of the VisualTasking Questionnaire of at least about 10%, at least about 20%, or atleast about 30%.

In some embodiments, the administration of the compound of formula Iresults in reduced ocular hyperemia (redness of the eye), compared toplacebo. In particular embodiments, the administration of the compoundof formula I results in reduced grade 1, grade 2, grade 3, or grade 4hyperemia compared to placebo. In some embodiments, the administrationresults in a reduction in ocular hyperemia score of at least about 1, atleast about 2, at least about 3, at least about 4, or at least about 5,on the McMonnies scale.

Thus, in some embodiments, the present invention relates to a method oftreating or reducing ocular hyperemia in a subject in need thereof,comprising administering to the subject an effective amount of compoundof formula I, or a salt, solvate, polymorph, or co-crystal thereof. Insome embodiments, the administration of compound I results in areduction in ocular hyperemia of at least 1, at least 2, at least 3, atleast 4, or at least 5 on the McMonnies scale. In some embodiments, theinvention provides for the use of the compound of formula I, or apharmaceutically acceptable salt, solvate, or co-crystal thereof, in thetreatment or reduction of ocular hyperemia. In some embodiments, theinvention provides for the administration of the compound of formula Ito a subject in need thereof in a ophthalmically compatible formulationat a concentration of about 0.5% w/v to about 3.5% w/v. In someembodiments, concentrations for administration range from about 0.5% toabout 3.5% w/v, about 0.5% to about 2.5% w/v, about 0.5% to about 1.5%w/v, about 0.5% to about 3.0% w/v, about 1.0% to about 2.5% w/v, about1.5% to about 3.0% w/v, about 0.5% to about 2.5% w/v. In particularembodiments, the concentration of the compound of formula I in aformulation for topical use is about 0.5% w/v, about 1.0% w/v, about1.5% w/v, about 2.0% w/v, about 2.5% w/v, about 3.0% w/v, or about 3.5%w/v. In some embodiments, the dose per administration per eye is fromabout 0.15 to about 1.15 mg, or about 0.15 mg, 0.2 mg, about 0.25 mg,0.3 mg, about 0.35 mg, about 0.4 mg, about 0.45 mg, about 0.5 mg, about0.55 mg, about 0.6 mg, about 0.65 mg, about 0.7 mg, about 0.75 mg, about0.8 mg, about 0.85 mg, about 0.9 mg, about 0.95 mg, about 1.0 mg, about1.05 mg, about 1.1 mg, or about 1.15 mg. In some embodiments, the doseper administration per eye is about 0.18 mg, about 0.37 mg, about 0.55mg, about 0.74 mg, or about 0.92 mg. In some embodiments, the totaldaily dose per eye is about 0.5 to about 3.5 mg, or about 0.5 mg, about1.0 mg, about 1.5 mg, about 2.0 mg, about 2.5 mg, about 3.0 mg, or about3.5 mg. In some embodiments, the compound of formula I is administeredto the subject one to six times a day, e.g., one, two, three, or fourtimes a day. In some embodiments, the compound of formula I isadministered to the subject for a period of at least about one month, atleast about two months, or at least about three months. In particularembodiments, the compound of formula I is administered in a formulationdescribed herein.

In some embodiments, the ocular hyperemia is associated with one or moreof dry eye disease, Sjogren's Syndrome, conjunctivitis (includingkeratoconjuctivitis, vernal keratoconjunctivitis, allergicconjunctivitis), Map-Dot-Fingerprint Dystrophy, acanthamoeba,fibromyalgia, Meibomian gland dysfunction, thyroid eye disease, rosacea,ptosis, keratoconus, ocular pain syndrome, Steven-Johnson's syndrome,corneal epitheliopathies, corneal neuropathies (including LASIK inducedcorneal neuropathies), corneal dystrophies (including recurrent cornealdystrophies), epithelial basement membrane dystrophy, corneal erosionsor abrasions (including recurrent corneal erosions or abrasions), ocularsurface diseases, blepharitis, graft vs host disease, meibomitis,glaucoma, conjunctivochalasis, keratopathis (including herpetickeratopathy, filamentary keratopathy, band or bullous keratopathy,exposure keratopathy), keratitis (including herpes simplex viruskeratitis), iritis, episclentis, corneal surgery, multiple sclerosis,trichiasis, pterygium, neuralgia, xerophthalmia, or patients recoveringfrom neurotrophic keratitis. In particular embodiments, the ocularhyperemia is associated with dry eye disease. In some embodiments of themethods described herein, the ocular hyperemia persists for at leastthree months after photorefractive keratectomy (PRK) surgery orlaser-assisted in situ keratomileusis (LASIK) surgery.

In some embodiments, the ocular surface pain or chronic ocular surfacepain is associated with dry eye disease. In some embodiments, theadministration of the compound of formula I results in a decrease in thesymptoms of dry eye disease. Dry eye disease is generally understood tobe a complex, multifactorial condition characterized by inflammation ofthe ocular surface and lacrimal glands and reductions in the qualityand/or quantity of tears. It is believed that up to 30% of dry eyedisease patients suffer from ocular surface pain that may be chronic,i.e., lasting at least 12 weeks or three months. Thus, in someembodiments, the invention results in a decrease of at least about 10%in the symptoms of dry eye disease, including one or more of oculardryness, ocular discomfort, ocular hyperemia, ocular burning orstinging, grittiness or foreign body sensation, or photophobia.

In some embodiments, the invention relates to a method of treating dryeye disease in a subject in need thereof, comprising administering tothe subject an effective amount of compound of formula I, or a salt,solvate, polymorph, or co-crystal thereof. In some embodiments, theinvention relates to a method of treating dry eye disease in a subjectin need thereof, comprising administering to the subject an effectiveamount of compound of formula I, or a salt, solvate, polymorph, orco-crystal thereof, wherein the compound of formula I is safe foradministration over a period of at least 2 months, at least 3 months, atleast 4 months, or at least 5 months. In particular embodiments, theinvention provides for the use of the compound of formula I, or apharmaceutically acceptable salt, solvate, or co-crystal thereof, in thetreatment of dry eye disease. In some embodiments, the invention resultsin a decrease of at least about 10%, at least about 15%, at least about20%, or at least about 30% in the symptoms of dry eye disease, includingone or more of ocular dryness, ocular discomfort, ocular hyperemia,ocular burning or stinging, grittiness or foreign body sensation, orphotophobia. In some embodiments, the invention provides for theadministration of the compound of formula I to a subject in need thereofin a ophthalmically compatible formulation at a concentration of about0.5% w/v to about 3.5% w/v. In some embodiments, concentrations foradministration range from about 0.5% to about 3.5% w/v, about 0.5% toabout 2.5% w/v, about 0.5% to about 1.5% w/v, about 0.5% to about 3.0%w/v, about 1.0% to about 2.5% w/v, about 1.5% to about 3.0% w/v, about0.5% to about 2.5% w/v. In particular embodiments, the concentration ofthe compound of formula I in a formulation for topical use is about 0.5%w/v, about 1.0% w/v, about 1.5% w/v, about 2.0% w/v, about 2.5% w/v,about 3.0% w/v, or about 3.5% w/v. In some embodiments, the dose peradministration per eye is from about 0.15 to about 1.15 mg, or about0.15 mg, 0.2 mg, about 0.25 mg, 0.3 mg, about 0.35 mg, about 0.4 mg,about 0.45 mg, about 0.5 mg, about 0.55 mg, about 0.6 mg, about 0.65 mg,about 0.7 mg, about 0.75 mg, about 0.8 mg, about 0.85 mg, about 0.9 mg,about 0.95 mg, about 1.0 mg, about 1.05 mg, about 1.1 mg, or about 1.15mg. In some embodiments, the dose per administration per eye is about0.18 mg, about 0.37 mg, about 0.55 mg, about 0.74 mg, or about 0.92 mg.In some embodiments, the total daily dose per eye is about 0.5 to about3.5 mg, or about 0.5 mg, about 1.0 mg, about 1.5 mg, about 2.0 mg, about2.5 mg, about 3.0 mg, or about 3.5 mg. In some embodiments, the compoundof formula I is administered to the subject one to six times a day,e.g., one, two, three, or four times a day. In some embodiments, thecompound of formula I is administered to the subject for a period of atleast about one month, at least about two months, or at least aboutthree months. In some embodiments, the compound of formula I isadministered as a formulation described herein.

In some embodiments of the methods described herein, the administrationof the compound of formula I does not result in a change (e.g., of lessthan 5% difference, less than 4% difference, or less than 3% difference)in one or more of best corrected visual acuity, slit-lamp biomicroscopy,dilated eye exam, blink rate, tear production, or intraocular pressurecompared to a placebo. In some embodiments of the methods describedherein, the administration of compound of formula I does not result in adelay in wound healing compared to a placebo in a patient in needthereof. Patient Population

In specific embodiments, a subject to be treated by methods providedherein suffers from an ocular surface disorder. Non-limiting examples ofocular surface disorders include chronic ocular surface pain (COSP), dryeye disease, Sjogren's Syndrome, conjunctivitis (includingkeratoconjuctivitis, vernal keratoconjunctivitis, allergicconjunctivitis), Map-Dot-Fingerprint Dystrophy, acanthamoeba,fibromyalgia, Meibomian gland dysfunction, thyroid eye disease, rosacea,ptosis, keratoconus, ocular pain syndrome, Steven-Johnson's syndrome,corneal epitheliopathies, corneal neuropathies (including LASIK inducedcorneal neuropathies), corneal dystrophies (including recurrent cornealdystrophies), epithelial basement membrane dystrophy, corneal erosionsor abrasions (including recurrent corneal erosions or abrasions), ocularsurface diseases, blepharitis, graft vs host disease, meibomitis,glaucoma, conjunctivochalasis, keratopathis (including herpetickeratopathy, filamentary keratopathy, band or bullous keratopathy,exposure keratopathy), keratitis (including herpes simplex viruskeratitis), iritis, episclentis, corneal surgery, multiple sclerosis,trichiasis, pterygium, neuralgia, xerophthalmia, or patients recoveringfrom neurotrophic keratitis. In some embodiments, the subject suffersfrom ocular pain persisting for at least three months afterphotorefractive keratectomy (PRK) surgery or laser-assisted in situkeratomileusis (LASIK) surgery.

In certain embodiments, methods provided herein is for treating, orreducing, ocular surface pain, such as acute ocular surface pain.

In certain embodiments, methods provided herein is for treating, orreducing, ocular surface pain, such as chronic ocular surface pain(COSP). In particular aspects, COSP is characterized as persistentocular surface pain (e.g., persistent severe ocular surface pain) thatcan distract from, or can interfere with, regular daily activities. Inspecific aspects, COSP can result in poor quality of life, and canpersist for at least 1 month, at least 2 months, at least 3 months, atleast 4 months, at least 5 months, or at least 6 months. In someaspects, COSP can persist for at least about 2 months or at least about3 months. In other aspects, COSP can persist for at least 3 months or atleast 4 months. In particular aspects, subject with COSP remainsymptomatic despite adherence to other therapies indicated for theirunderlying disease (e.g., an ocular surface disorder such as dry eyedisease or Sjogren's Syndrome).

In some embodiments, the subject to be treated suffers from ocularneuropathic pain (ONP). ONP is a spectrum of disorders of ocular painthat may be caused by damage or disease affecting the nerves, e.g.,corneal nerves. Symptoms of ONP may include one or more of eye pain,sensitivity to light, hyperalgesia or dysesthesia (abnormal sensations)such as a sensation of dryness, stinging, or foreign body, pain fromnormally non-painful stimuli (allodynia). Gabapentin and otherneuropathic pain medications may be used to blunt sensory nervestimulation or the perception of nerve stimulation.

In some embodiments, the subject to be treated suffers from exposurekeratopathy. EK is damage to the cornea that occurs primarily fromprolonged exposure of the ocular surface to the outside environment. EKcan lead to ulceration, microbial keratitis, and permanent vision lossfrom scarring. Patients at risk for EK include those who suffer fromconditions that interfere with the ability to protect the cornea; eitherby incomplete eyelid closure (e.g., lagophthalmos, proptosis, lidmalposition), inadequate blink reflex, inadequate blink rate (forexample, caused by a neurologic disease, e.g., Parkinson disease, aneuromuscular disease) and/or decreased protective lubrication of thecornea. Symptoms of EK include foreign body sensation, burning,increased tearing, and intermittent blurry vision (from an unstable tearfilm), pain and photophobia. Standard treatments include the use offrequent artificial tears with nightly lubricating ointment, punctalplugs.

In some embodiments, the subject to be treated suffers fromkeratoconjunctivitis. Keratoconjuctivitis is an inflammatory processthat involves both the conjunctiva and the cornea. Superficialinflammation of the cornea (keratitis) occurs commonly in associationwith viral and bacterial conjunctivitis, for example in adults. Thefollowing types of keratoconjuctivitis are distinguished based on thepotential cause of inflammation:

-   -   Keratoconjunctivitis sicca is cause by the inflammation due to        dryness;    -   Vernal keratoconjunctivitis (VKC) occurs seasonally, considered        to be due to allergens;    -   Atopic keratoconjunctivitis is one manifestation of atopy;    -   Epidemic keratoconjunctivitis or adenoviral keratoconjunctivitis        is caused by an adenovirus infection;    -   Infectious bovine keratoconjunctivitis (IBK) is a disease        affecting cattle caused by the bacteria Moraxella bovis;    -   Pink eye in sheep and goat is mostly caused by Chlamydophila        pecorum;    -   Superior limbic keratoconjunctivitis is thought to be caused by        mechanical trauma;    -   Keratoconjunctivitis photoelectrica (arc eye) means inflammation        caused by photoelectric UV light.

In some embodiments, the subject to be treated suffers from dry eye. Theterm “dry eye” as used herein, refers to inadequate tear productionand/or abnormal tear composition. Dry eye syndrome disease (DEDS), alsoknown as dry eye syndrome, keratoconjunctivitis sicca or keratitissicca, or tear dysfunction syndrome, or burning eye syndrome resultsfrom deficiency of any of the tear film layers. Dry eye is amultifactorial disease of the tears and ocular surface that results insymptoms of discomfort, visual disturbance, and tear ¬film instabilitywith potential damage to the ocular surface ocular surface characterizedby loss of homeostasis of the tear film, and accompanied by ocularsymptoms, in which tear film instability and hyperosmolarity, ocularsurface inflammation and damage, and neuro-sensory abnormalities playetiological roles (Craig J P, et al., The Ocular Surface 2017;15:276-83). It may be accompanied by increased osmolarity of the tearfilm and inflammation of the ocular surface. Dry eye disorder may rangefrom mild to moderate to severe forms. Symptoms of dry eye syndromedisease include gritty, foreign body sensations, burning, photophobia,and decreased visual acuity, tearing, stinging, itching, sandy or grittyfeeling, discharge, frequent blinking, mattering or caking of theeyelashes (usually worse upon waking), redness, blurry or fluctuatingvision (made worse when reading, computer, watching television, driving,or playing video games), light-sensitivity, eye pain and/or headache,heavy eye lids, eye fatigue. Causes of dry eye disease include, but arenot limited to, the following: idiopathic, congenital alacrima,xerophthalmia, lacrimal gland ablation, and sensory denervation;collagen vascular diseases, including rheumatoid arthritis, Wegener'sgranulomatosis, and systemic lupus erythematosus; Sjogren's Syndrome andautoimmune diseases associated with Sjogren's syndrome; abnormalities ofthe lipid tear layer caused by blepharitis or rosacea; abnormalities ofthe mucin tear layer caused by vitamin A deficiency; trachoma,diphtheric keratoconjunctivitis; mucocutaneous disorders; aging;menopause; and diabetes. Dry eye signs and/or symptoms as defined hereinmay also be provoked by other circumstances, including but not limitedto the following: prolonged visual tasking; working on a computer; beingin a dry environment; warm or cold wind or air flow; seasonal changes;ocular irritation; contact lenses, LASIK and other refractive surgeries;fatigue; and medications such as isotretinoin, sedatives, diuretics,tricyclic antidepressants, antihypertensives, oral contraceptives,antihistamines, nasal decongestants, beta-blockers, phenothiazines,atropine, and pain relieving opiates such as morphine.

Diagnostic testing for dry eye includes evaluation of cornea sensation(corneal hyperesthesia and/or reduced sensation may be present in severeand chronic dry eye disease) using, for example, a cotton tip applicatoror more precisely with a Cochet-Bonnet esthesiometer; measuring tearbreak up time using, for example, a fluorescein-impregnated strip wetwith non-preserved saline solution or more objective computerizedmethods without the need for fluorescein instillation; performing ocularsurface staining, e.g., fluorescein sodium, rose bengal, lissaminegreen; performing Schirmer test (relatively insensitive for patientswith mild dry eye), testing delayed tear clearance; tear meniscusheight; measuring level of MMP-9 (MMP-9 has been shown to be elevated inthe tears of patient with dry eye disease, and levels correlate withexamination findings in patients with moderate to severe dry eye);measuring tear osmolarity and tear film interferometry; performing Sjotest (detection of SS-A (anti-Ro) and SS-B (anti-La) autoantibodies inserum, salivary gland protein 1 (SP-1), carbonic anhydrase 6 (CA6), andparotid secretory protein (PSP). SP-1, CA6, and PSP).

Artificial tears, lubricating ointments, corticosteroids (e.g.,loteprednol 0.5% eyedrops four times a day) are used as an initialtreatment. Prescription medicines include cyclosporine, lifitegrast,diquafosol, rebamepide, corticosteroids (e.g., loteprednol 0.5% eyedropsfour times a day).

The term “tear film dysfunction” refers to a state when the tear filmbreaks down in different places on the cornea and conjunctiva, leadingnot only to symptoms of irritation, but also to unstable andintermittently changing vision. For example, dry eye syndrome disease ischaracterized by tear film dysfunction. The symptoms of tear filmdysfunction include tearing, burning, stinging, itching, sandy or grittyfeeling, scratchy or foreign-body sensation, discharge, frequentblinking, mattering or caking of the eyelashes (usually worse uponwaking), redness, blurry or fluctuating vision (made worse when reading,computer, watching television, driving, or playing video games),light-sensitivity, eye pain and/or headache, heavy eye lids, eyefatigue.

Adenoviral keratoconjunctivitis, also known as Keratoconjunctivitisepidemica is a common and highly contagious viral infection of the eye.The clinical course of Adenoviral keratoconjunctivitis is divided intoan acute phase with conjunctival inflammation of varying intensity withor without corneal involvement and a chronic phase with cornealopacities.

Vernal keratoconjunctivitis (VKC) is an atopic condition of the externalocular surface characterized by symptoms consisting of severe itching,photophobia, foreign body sensation, mucous discharge (often describedas “ropy”), blepharospasm, and blurring of vision (Buckley, R. J., IntOphthalmol Clin, 1988 28(4): p. 303-8; Kumar, S., Acta Ophthalmologica,2009. 87(2): p. 133-147). It is typically bilateral but may beasymmetric in nature. It characteristically affects young males in hotdry climates in a seasonal manner; in 23% of patients may have aperennial form (Kumar, S., Acta Ophthalmologica, 2009. 87(2): p.133-147; Bonini, S., et al., Ophthalmology, 2000. 107(6): p. 1157-63).

The signs of VKC can be divided into conjunctival, limbal and cornealsigns:

-   -   Conjunctival signs include diffuse conjunctival injection and        upper tarsal giant papillae that are discrete >1 mm in diameter;    -   Limbal signs include thickening and opacification of the limbal        conjunctiva as well as gelatinous appearing and sometime        confluent limbal papillae. Peri-limbal Horner-Trantas dots are        focal white limbal dots consisting of degenerated epithelial        cells and eosinophils (Buckley, R. J., Int Ophthalmol        Clin, 1988. 28(4): p. 303-8);    -   Corneal signs vary according to the severity of the disease        process and include macro-erosions, cornal ulcers and scars        (Buckley, R. J., Int Ophthalmol Clin, 1988. 28(4): p. 303-8).

Active VKC patients (defined as moderate to severe ocular discomfortincluding photophobia, papillae on the upper tarsal conjunctiva, orlimbal Horner-Trantas dots clearly recognizable at the time of theexamination) showed significantly increased symptoms and signs of ocularsurface disease. Inactive VKC patients (defined as no symptoms or milddiscomfort, and absence of corneal abnormalities at the time of theexamination) showed increased photophobia, conjunctival lissamine greenstaining and Schirmer test values, and reduced fluorescein break-up time(BUT) and corneal sensitivity. This syndrome seems to affect the ocularsurface in all phases (active and quiescent), determining abnormalitiesin tear film stability, epithelial cells integrity, and corneal nervesfunction (Villani E. et al., Medicine (Baltimore). 2015 October; 94(42):e1648).

The following factors are thought to play a role in VKC: IgE mediatereaction via mast cell release; activated eosinophils, mononuclear cellsand neutrophils as well as the CD4 T-helper-2 driven type IVhypersensitivity with immunomodulators such as IL-4, IL-5, and bFGF(Buckley, R. J., Int Ophthalmol Clin, 1988. 28(4): p. 303-8; Kumar, S.,Acta Ophthalmologica, 2009. 87(2): p. 133-147; La Rosa, M., et al., ItalJ Pediatr, 2013. 39: p. 18).

Treatment consists of cool compresses and lid scrubs, saline eyedrops,which may help to relieve symptoms, along with topical antihistamines,nonsteroidal anti-inflammatory drugs or corticosteroids, e.g.,low-absorptions corticosteroids (fluoromethelone, loteprednol,remexolone, etc.), optical mast cell stabilizers (cromolyn sodium,nedocromil sodium, and lodoxamide), topical cyclosporin-A, ortacrolimus. See e.g., Oray, M. and E. Toker, Cornea, 2013. 32(8): p.1149-54: Vichyanond, P. and P. Kosrirukvongs, Curr Allergy Asthma Rep,2013. 13(3): p. 308-14; Barot, R K et al., J Clin Diagn Res. 2016 June;10(6):NC05-9; Wan Q et al., Ophthalmic Res. 2018; 59(3):126-134.

Atopic keratoconjunctivitis (AKC) typically has an older age of onset inthe 2nd to 5th decade, as opposed to onset prior to age 10 with VKC.Conjunctival involvement is classically on the upper tarsus in VKC andon the lower tarsus in AKC. AKC is typically more chronic in nature andmore commonly results in scarring of the cornea and conjunctivalcicatrization.

Sjogren's Syndrome (Sjogren's syndrome associated with dry eye) is achronic inflammatory disorder characterized by exocrine glanddysfunction including the salivary and lacrimal glands that in manycases results in a severe dry eye. Primary symptoms are dry eyes(keratitis sicca or keratoconjunctivitis sicca) and dry mouth(xerostomia). Severe dry eyes can cause corneal pain, corneal scarring,ulceration, infection, and even perforation. The differential diagnosisincludes conditions such as adult blepharitis, dry eye disease, andjuvenile idiopathic arthritis uveitis, as well keratopathies, e.g.,superficial punctate, filamentary, neurotrophic, exposure). Treatment ofSjogren's syndrome is aimed at maintaining the integrity of the tearfilm through preservation, augmentation, and/or replacement of thedeficient tear secretion. Treatment of Sjogren's syndrome thus includesartificial tears and lubricating ointments; autologous serum eyedrops;oral omega-6 essential fatty acids; fluid-ventilated, gas permeablescleral lenses; topical corticosteroids; punctal occlusion to decreasetear drainage; a small lateral tarsorrhaphy; humidification of theenvironment; hydrophilic bandage lenses; bromhexine and 3-isobutyl1-methylxanthine (IBMX) (augmentation of tear production/secretion);agents to stimulate muscarinic receptors (pilocarpine and cevimeline);immunosuppressive agents, e.g., methotrexate, antimalarials,cyclophosphamide, leflunomide, or tumor necrosis factor (TNF), e.g.,infliximab, a monoclonal antibody to TNF-alpha; Cyclosporin A; thebandage contact lens.

Steven-Johnson's syndrome (SJS) is a dermatologic emergency or a type ofsevere skin reaction characterized by the presence of epidermal andmucosal bullous lesions involving less than 10% of the total bodysurface area. Early symptoms of SJS include fever and flu-like symptoms,which may precede or occur concurrently with the development of amacular rash involving the trunk and face. As the disease progresses,the macular rash coalesces, the involved areas develop bullae, and theepidermal layer eventually sloughs off. During the acute phase ofSJS-TEN, 80% of patients will have ocular involvement.

The constellation of high fever (>102.2), malaise, arthralgia, a macularrash involving the trunk, neck and face, and recent history of newmedication exposure or recently increased dosage of an existingmedication are indicators used for diagnosis of SJS. A skin biopsy of aneffected area can be performed for a confirmation of the diagnosis.Granulysin can be used as a marker for the diagnosis of SJS. Theconcentration of granulysin within bullous fluid correlates with theseverity of the acute phase of SJS (Chung W H, et al. Nat Med. 2008;14(12):1343-50).

Ocular manifestations in SJS include conjunctivitis, subconjunctivalhemorrhage, subconjunctival scarring, conjunctival membranes,conjunctival ulceration, superficial punctate epithelial erosions,epithelial defects, lid margin ulceration, lid margin keratinization,symblepharon, ankyloblepharon, trichiasis, anterior blepharitis, punctalauto-occlusion, meibomian gland disease, corneal opacification, dry eye,districhiasis, limbal stem cell failure, corneal vascularization. Eyetreatment in SJS consists of saline eyedrops, preservative-freeartificial tears and ointments to provide adequate lubrication andreduce epithelial injury. Patients with any corneal or conjunctivalepithelial defects are treated with prophylactic topical antibiotics,preferably a fourth generation fluoroquinolone. Patients having mild ormoderate ocular involvement (less than one-third lid margin involvement,conjunctival defects less than 1 cm at greatest diameter, and no cornealepithelial defects) are typically treated with topical moxifloxacin 0.5%four times a day, cyclosporine 0.05% twice daily, and topical steroids(prednisolone acetate 1% four to eight times a day or dexamethasone 0.1%twice daily). Patients having severe or extremely severe ocularinvolvement (greater than one-third lid margin involvement, conjunctivaldefects greater than 1 cm, and corneal epithelial defects) undergo anamniotic membrane (AM) grafting in addition to the treatments listedabove.

In some embodiments, the subject to be treated suffers from cornealepitheliopathy. Corneal epitheliopathy is a disease involving cornealepithelium, e.g., manifested in altered corneal epithelial barrierfunction.

In some embodiments, the subject to be treated suffers from cornealneuropathy or corneal neuralgia. Corneal neuropathy or corneal neuralgiais a disorder associated with corneal pain caused by the damaged nervefibers in the cornea, the sensory fibers. One of the examples of cornealneuropathy is a LASIK induced corneal neuropathy. Corneal neuropathygenerally could be identified and diagnosed through dry eyeinvestigations. Though the causes and risk factors are unclear yet,patients with dry eye-like symptoms, increased corneal sensitivity andchanges of corneal nerve morphology, but no signs of dryness may sufferfrom corneal neuropathy.

In some embodiments, the subject to be treated suffers from ocularsurface disease or disorder. The term “ocular surface diseases” or“ocular surface disorders” encompasses disease entities as well asrelated symptoms that result from a variety of abnormalities, includingabnormal lid anatomy or function, abnormal or altered tear production orcomposition, and related subclinical signs. Many diseases can causeocular surface disorders. Patients with ocular surface disorders mayexhibit clinical signs common to several diseases, and include chronicpunctate keratopathy, filamentary keratopathy, recurrent cornealerosion, bacterial conjunctivitis, culture-negative conjunctivitis,cicatrising (scarring) conjunctivitis, persistent epithelial defect,infectious keratitis, corneal melt and ocular surface failure. The mostcommon ocular surface disorders stem from tear-film abnormalities and/orlid-gland dysfunction (“blepharitis”).

In some embodiments, the subject to be treated suffers from neurotrophickeratitis or neurotrophic keratopathy. Neurotrophic keratitis orneurotrophic keratopathy (NK) is a corneal degenerative diseasecharacterized by a reduction or absence of corneal sensitivity. In NK,corneal innervation by trigeminal nerve is impaired. Since cornealsensory innervation is impaired in NK, patients do not commonly complainof ocular surface symptoms. However, blurred vision can be reported dueto irregular epithelium or epithelial defects (PED), scarring, or edema.NK is usually graded in three different stages in accordance to the“Mackie classification”. Stage II NK is defined by a recurrent orpersistent epithelial defects, most commonly in the superior half of thecornea. One of the treatments that may be used in Stage II NK includestopical Nerve Growth Factor. Patients typically experience pain duringtreatment with NGF due to reforming of the nerves.

In some embodiments, the subject to be treated suffers from blepharitis.Blepharitis is an inflammatory condition of the eyelid margin, which canlead to permanent alterations in the eyelid margin or vision loss fromsuperficial keratopathy, corneal neovascularization, and ulceration.According to anatomic location, blepharitis can be divided into anteriorand posterior. Anterior blepharitis affects the eyelid skin, base of theeyelashes, and the eyelash follicles and includes the traditionalclassifications of staphylococcal and seborrheic blepharitis. Posteriorblepharitis affects the meibomian glands and gland orifices, the primarycause being meibomian gland dysfunction. Symptoms of chronic blepharitismay include redness, burning sensation, irritation, tearing, eyelidcrusting and sticking, and visual problems such as photophobia andblurred vision. Long-term management of symptoms may include dailyeyelid cleansing routines and the use of therapeutic agents that reduceinfection and inflammation. Treatment includes topical or systemicantibiotics e.g., bacitracin or erythromycin; oral antibiotics, e.g.,tetracyclines (tetracycline, doxycycline, minocycline) or macrolides(erythromycin, azithromycin); topical steroids, e.g., corticosteroid,e.g., loteprednol etabonate, fluorometholone; topical combinations of anantibiotic and corticosteroid such as tobramycin/dexamethasone ortobramycin/loteprednol; topical cyclosporine 0.05%.

In some embodiments, the subject to be treated suffers from Meibomiangland dysfunction. The meibomian gland is a holocrine type of exocrinegland, at the rim of the eyelid inside the tarsal plate, responsible forthe supply of meibum, an oily substance that prevents evaporation of theeye's tear film. Meibomian gland dysfunction (MGD), also known asmeibomitis, posterior blepharitis or inflammation of the meibomianglands, is a chronic, diffuse abnormality of the meibomian glands,commonly characterized by terminal duct obstruction and/orqualitative/quantitative changes in the glandular secretion (Nelson J D,et al., Invest Ophthalmol Vis Sci 2011; 52:1930-7). It may result inalteration of the tear film, symptoms of eye irritation, clinicallyapparent inflammation, and ocular surface disease. MGD often causes dryeye, and may contribute to blepharitis. In some cases topical steroidsand topical/oral antibiotics are also prescribed reduce inflammation.Intense pulsed light (IPL) treatments or other mechanical treatmentsthat apply heat and pressure to express the glands (eg, LipiFlow) havealso been shown to reduce inflammation and improve the gland function inpatients.

In some embodiments, the subject to be treated suffers fromgraft-versus-host disease. Graft-versus-host disease (GVHD) is aninflammatory disease that is unique to allogeneic transplantation. It isan attack by transplanted leukocytes against the recipient's tissuesthat can occur even if the donor and recipient are HLA-identical. Acutegraft-versus-host disease typically occurs in the first 3 months aftertransplantation and may involve the skin, intestine, or the liver.Corticosteroids such as prednisone are a standard treatment. Chronicgraft-versus-host disease may also develop after allogeneic transplantand is the major source of late complications. In addition toinflammation, chronic graft-versus-host disease may lead to thedevelopment of fibrosis, or scar tissue, similar to scleroderma or otherautoimmune diseases and may cause functional disability, and the needfor prolonged immunosuppressive therapy.

In some embodiments, the subject to be treated suffers from ocular graftversus host disease. GVHD occurs in patients who have undergoneallogenic hematological stem cell transplantation. It can occur inpatients who have acute or chronic GVHD, though it is more common inpatients with the chronic form. Approximately 40-90% of patients withchronic GVHD will develop ocular symptoms. Ocular manifestations caninclude moderate to severe keratoconjunctivitis sicca, bilateralmarginal keratitis, anterior uveitis, corneal ulceration orneovascularization. Treatment includes topical lubricants includingpreservative free artificial tears, autologous serum tears and othertopical and systemic immunosuppressive treatments; systemic steroids;topical cyclosporine 0.5%.

EXAMPLES

The following examples are included to demonstrate non-limitingembodiments of the present invention. Those of skill in the art willappreciate that changes to the specific embodiments described herein canbe made and still obtain a like result without departing from the spiritand scope of the invention.

Example 1. Solubility of Compound I

As noted above, compound I is sparingly soluble in various media. Thesolubility of compound I in various solvents is shown in Table 1.

TABLE 1 Solubility of compound I in Solvents Solvent Solubility Water0.01 Ethanol 0.79 Polyethylene Glycol 200 1.62 Polyethylene Glycol 3002.04 Polyethylene Glycol 400 1.66 Propylene Glycol 1.68 Isosorbide 0.34Propylene Carbonate 0.40 Hexylene Glycol 0.28 Isopropanol 0.33 IsopropylMyrisate 0.09 Diisopropyl Adipate 0.16 Oleyl Alcohol 0.12 Mineral Oil0.03 Medium-Chain Triglycerides (Miglyol 812) 0.10

Example 2. Exploratory Formulations for Stability Testing

Based on the limited solubility of compound I in various solvents and inwater, a suspension formulation was explored for development. Thefollowing five formulations were prepared and tested for stability.Formulations were stored in crimp-sealed glass vials at room temperatureand at 40° C. At 6, 8 and 12 weeks, samples were evaluated for compoundI stability.

TABLE 2 Compound I Formulations for Exploratory Stability Percent w/wFID FID FID FID Lot Component 121522 121511 121512 121513^(a) 17307:087BCompound I 0.5 0.5 0.5 0.00135 0.5 Tyloxapol 0.1 0.1 0.1 0.1 0.1Poloxamer 407 17.5 — — — — Hypromellose — 0.5 0.5 — — CarbomerHomopolymer Type B — — — — 0.4 Polyethylene Glycol 6000 2 — — — —Polyethylene Glycol 400 — — 7 7 — Benzalkonium Chloride 0.02 0.02 0.020.02 0.02 Edetate Disodium 0.02 0.02 0.02 0.02 0.02 Boric Acid 0.3 0.30.3 0.3 0.3 Mannitol 0.3 4.5 0.3 0.3 4.5 Monobasic Sodium 0.13 0.13 0.130.13 0.13 Phosphate monohydrate Dibasic sodium 0.01 0.01 0.01 0.01 0.01phosphate anhydrous Sodium Hydroxide qs pH 6 qs pH 6 qs pH 6 qs pH 6 qspH 6 Hydrochloric Acid qs pH 6 qs pH 6 qs pH 6 qs pH 6 qs pH 6 PurifiedWater qs 100 qs 100 qs 100 qs 100 qs 100 ^(a)FID 121513 is the filteredsupernatant of FID 121512 and is a solution. The other four formulationsin the table are suspensions.

The formulations prepared in Table 2 were assayed using an exploratoryUPLC method developed for the assay of compound I as shown in Table 3.

TABLE 3 Exploratory Compound I UPLC Method Column ACQUITY UPLC BEHShield RP18, 1.7 μm, 2.1 × 100 mm Column temperature 65° C. UV detectorwavelength  254 nm Injection volume 2 μL Mobile phase A 0.1% Formic AcidB Acetonitrile Tim Flow Rate Gradient (min (mL/min % A % B Curve 0 0.870 30 1.0 0.8 30 70 6 1.5 0.8 20 80 6 2 0.8 70 30 6 Run time 3 Relativeretention 0.9

Results from the testing for compound I stability at room temperatureand 40° C. are shown in FIG. 1A and FIG. 1B, respectively. In FIG. 1A,the formulation with percent of compound I at about 70% of original at12 weeks is the formulation of compound I as a solution. In FIG. 1B, theformulation with percent of compound I at about 20% of original at 12weeks is the formulation of compound I as a solution. As seen in FIGS.1A and 1B compound I remains in stable form in the suspensions, withoutsignificant degradation even at 12 weeks at 40° C. In contrast, the partof compound I in solution in the supernatant (FID121513) degrades toabout 20% of initial amount after 12 weeks at 40° C.

At the end of the study, compound I was recovered from the foursuspensions and evaluated for changes in crystalline form by x-raypowder diffraction. The x-ray diffraction patterns of compound Irecovered from the 12-week stability samples and that of compound Istored at room temperature (control) are presented in FIG. 2 . As seenin FIG. 2 , compound I in each of the four exploratory formulationsmaintains its polymorphic form even after storage at room temperature or40° C. for 12 weeks.

In addition, the suspension FID 121744 (shown below in Table 6) and a10% compound I slurry in 1% tyloxapol were subjected to heatsterilization 171° C. for 1 hour. The X-ray diffraction pattern ofcompound I after sterilization was identical to an untreated sample.

Example 3. Viscosity Range Finding Studies of Compound I Suspensions

Various suspension formulations of compound I were evaluated forviscosity and settling of compound I. A series of unpreserved compound Isuspensions were prepared containing varying amounts of carbopol(Carbomer homopolymer Type B) and varying amounts of sodium chloride.The compositions of the formulations are shown in Table 4.

TABLE 4 Compositions of compound I unpreserved suspensions of compound Ifor viscosity range finding studies FID: 121845 121724 121846 121847121848 Component Percent w/w Compound I 0.5 0.5 0.5 0.5 0.5 Tyloxapol0.05 0.05 0.05 0.05 0.05 Carbopol 0.1 0.2 0.3 0.4 0.5 974P ^(a) Glycerin2.0 2.0 2.0 2.0 2.0 Sodium — — — — — Chloride Tro- qs pH 7.4 qs pH 7.4qs pH 7.4 qs pH 7.4 qs pH 7.4 methamine Hydrochloric qs pH 7.4 qs pH 7.4qs pH 7.4 qs pH 7.4 qs pH 7.4 Acid Purified qs 100 qs 100 qs 100 qs 100qs 100 Water ^(a) Carbomer homopolymer Type B

The suspensions were evaluated visually for uniformity. The suspensionwhich contained only 0.1% Carbomer, FID 121845, was not uniform andtherefore was not evaluated further. Sodium chloride is known to reducethe viscosity of Carbomer-containing suspensions, so sodium chloride wasadded to aliquots of the remaining four suspensions to obtain a total of28 suspensions with concentrations of 0.2, 0.3, 0.4 and 0.5% Carbomerand 0, 0.05, 0.1, 0.15, 0.2, 0.25 and 0.3% sodium chloride.

Results from the viscosity testing of 28 suspensions are shown in Table5.

TABLE 5 Viscosities (cP) of compound I unpreserved suspensions fromTable 4 (CP52, 60 RPM @ 25° C.) FID Sodium chloride (% w/w) (% carbomer)0 0.05 0.1 0.15 0.2 0.25 0.3 121724 (0.2) Out 64.5 23.9 14.6 10.4 7.597.44 of range 121846 (0.3) Out Out 101.2 59.8 38.0 25.6 18.9 of range ofrange 121847 (0.4) Out Out Out Out 121.1 84.0 63.7 of range of range ofrange of range 121848 (0.5) Out Out Out Out Out Out 150.8 of range ofrange of range of range of range of range

As seen from Table 5, the relationship between the amounts of carbomerand sodium chloride affect the viscosity of the suspension.

A further set of unpreserved suspensions of compound I containingcarbopol as a suspending agent were prepared for viscosity range findingand settling studies. The compositions, their viscosities and settlingtimes of 10 ml of suspension after 6 months are shown in Table 6.

TABLE 6 Unpreserved suspensions of compound I and their viscosities andsettling properties FID: 121744 121896 121897 121898 121899 ComponentPercent w/w Compound I 0.5 0.5 0.5 0.5 0.5 Tyloxapol 0.05 0.05 0.05 0.050.05 Carbopol 974P ^(a) 0.2 0.2 0.2 0.2 0.2 Glycerin 2.0 2.0 2.0 2.0 2.0Sodium Chloride 0.05 0.06 0.07 0.08 0.09 Tromethamine qs pH 7.4 qs pH7.4 qs pH 7.4 qs pH 7.4 qs pH 7.4 Hydrochloric Acid qs pH 7.4 qs pH 7.4qs pH 7.4 qs pH 7.4 qs pH 7.4 Purified Water qs 100 qs 100 qs 100 qs 100qs 100 Viscosity (cP) 153.4 113.2 84.8 66.4 49.8 Settling, ml (%) <0.2(<2%) <0.2 (<2%) <0.2 (<2%) — <0.2 (<2%) ^(a) Carbomer Homopolymer TypeB

A further series of preserved suspensions containing hypromellose as thesuspending agent were prepared and their viscosities measured. Thesettling properties were measured as follows: well mixed aliquots of thesuspensions were filled into 10 ml glass graduated cylinders. Thegraduated cylinders were sealed with ground glass stoppers and Parafilm®and allowed to sit undisturbed for six months at room temperature. Thesettled suspension was estimated by visual inspection. The compositions,their viscosities and settling times of 10 ml of suspension after 6months are shown in Table 7.

TABLE 7 Preserved suspensions of compound I and their viscosities andsettling properties FID: 121801 121901 121902 121903 Component Percentw/w Compound I 0.5 0.5 0.5 0.5 Tyloxapol 0.05 0.05 0.05 0.05Hypromellose 0.5 0.6 0.7 0.8 Boric Acid 0.3 0.3 0.3 0.3 Mannitol 0.3 0.30.3 0.3 Propylene Glycol 1.5 1.5 1.5 1.5 Polyquaternium-1 0.001 0.0010.001 0.001 Sodium Hydroxide qs pH 7.4 qs pH 7.4 qs pH 7.4 qs pH 7.4Hydrochloric Acid qs pH 7.4 qs pH 7.4 qs pH 7.4 qs pH 7.4 Purified Waterqs 100 qs 100 qs 100 qs 100 Viscosity (cP) 20.0. 32.0 50.2 76.4Settling, ml (%) 0.6 (6%) 0.4 (4%) 0.2 (2%) —

Based on the results shown in Table 6 and Table 7, compound Isuspensions that are preserved or unpreserved and that have eithercarbomer or hypromellose as a suspending agent were able to achieveacceptable viscosities and settling properties. As described supra,acceptable viscosities for the formulations of the present disclosureare in the range of about 20-200 cPs, measured using a CP52 spindle at60 rpm and at room temperature.

Example 4. pH Dependent Stability Screening of Compound I Suspensions

In order to test the pH stability of compound I suspensions, theformulations listed in Table 8 below were prepared. All samples contain0.1% compound I in order to keep drug concentration consistent. As thisamount of drug was not soluble in any of the samples, 0.1% tyloxapol wasused as a surfactant to aid in re-suspending the material in thesamples. All samples (except the TRIS sample) contain an equivalentamount of phosphate buffer. All samples were placed in 20 mL glass vialsand put on condition at 60° C. At each assay time point, samples wereallowed to equilibrate to room temperature, vortexed to resuspend thedrug, and samples were diluted to concentrations that fell within thelinear range of the standard curve.

TABLE 8 Compound I stability sample compositions pH 5 pH 6 pH 7 pH 8Carbopol HPMC TRIS Component Percent w/w Compound I 0.1 0.1 0.1 0.1 0.10.1 0.1 Tyloxapol 0.1 0.1 0.1 0.1 0.1 0.1 0.1 Hypromellose — — — — — 0.5— Carbopol — — — — 0.4 — — Monobasic 0.13 0.13 0.13 0.13 0.13 0.13 —sodium phosphate Dibasic sodium 0.01 0.01 0.01 0.01 0.01 0.01 —phosphate Tromethamine — — — — — — 0.03 Sodium hydroxide pH 5.0 pH 6.0pH 7.0 pH 8.0 pH 7.4 pH 7.4 pH 7.4 Hydrochloric acid pH 5.0 pH 6.0 pH7.0 pH 8.0 pH 7.4 pH 7.4 pH 7.4 Purified water Qs 100 Qs 100 Qs 100 Qs100 Qs 100 Qs 100 Qs 100

The amount of compound I in the stability samples was tested at 0, 4, 7,and 11 days by high performance liquid chromatography (HPMC), using aWaters XBridge Shield RP18 Column (3.5 μm, 3.0×150 mm, 30° C.), using agradient of mobile phase A 0.1% TFA in water and mobile phase B 0.1% TFAin acetonitrile, with an injection volume of 10 μl and a flow rate of0.8 ml/min. In addition, the major degradant appearing at a relativeretention time (RRT) of 1.23 was tracked. Results from the stabilitytesting are shown in Table 9.

TABLE 9 Stability of compound I and degradant growth at 60° C. CompoundI stability % Degradant @ (% initial) RRT 1.23 Condition Initial Day 4Day 7 Day 11 Initial Day 4 Day 7 Day 11 pH 5 100 95.5 91.2 85.6 0 2.32.8 3.9 pH 6 100 97.5 95.3 93.0 0 1.2 2.1 2.2 pH 7 100 95.7 92.3 87.8 02.0 2.2 4.0 pH 8 100 79.9 63.4 40.9 0 5.7 13.5 29.1 Carbopol 100 93.286.2 75.4 0 3.3 6.5 11.0 (pH 7.4) HPMC 100 94.4 89.8 83.3 0 2.9 5.1 8.2(pH 7.4) TRIS 100 98.6 96.2 94.4 0 1.1 2.0 2.6 (pH 7.4)

The major degradant shown in Table 9 was observed to crystallize out toform large crystals as its concentration increases. The followingformulations shown in Table 10 were therefore prepared and analyzed fordegradant formation.

TABLE 10 Compound I stability sample compositions 5% 5% SBE- 0.1% CaCl₂/0.5% 2% PVP HPβCD CD CaCl₂ HPMC HPMC 1% TRIS Component Percent w/vCompound I 0.1 0.1 0.1 0.1 0.1 0.1 0.1 Tyloxapol 0.1 0.1 0.1 0.1 0.1 0.10.1 Povidone K30 2 — — — — — — Hypromellose — — — — 0.5 0.5 — HP-β- — 5— — — — — cyclodextrin Sulfobutyl- — — 5 — — — — cyclodextrin Calcium —— — 0.1 0.1 — — chloride Sodium citrate 0.05 0.05 0.05 0.05 0.05 0.050.05 Tromethamine pH 6.0 pH 6.0 pH 6.0 pH 6.0 pH 6.0 pH 6.0 1.0 HCl pH6.0 pH 6.0 pH 6.0 pH 6.0 pH 6.0 pH 6.0 pH 6.0 Purified water Qs 100 Qs100 Qs 100 Qs 100 Qs 100 Qs 100 Qs 100

Data on the stability of compound I in compositions shown in Table 10 isshown below in Table 11.

TABLE 11 Stability of Compound I in Suspension at 40° C. and 60° C. %Initial of % Initial of 0.1% compound I at 40° C. compound I at 60° C.Compound I Initial Day 15 Day 35 Day 56 Initial Day 7 Day 15 2% PVP 10098.5 97.0 95.4 100 94.5 87.6 5% HP-β-CD 100 97.3 95.2 92.9 100 91.4 82.05% SBE-CD 100 92.4 87.1 84.0 100 83.3 66.9 0.1% CaCl₂ 100 98.4 97.6 96.3100 94.7 87.3 CaCl₂/HPMC 100 97.7 97.0 94.7 100 93.4 87.4 0.5% HPMC 10097.7 96.4 95.7 100 93.1 89.9 1% TRIS 100 97.4 95.9 93.7 100 89.8 79.5

Data on the degradant growth is shown in Table 12.

TABLE 12 Compound I degradant growth in suspension at 40° C. and 60° C.% Deg @ % Deg @ 0.1% RRT 1.23 at 40° C. RRT 1.23 at 60° C. Compound IInitial Day 15 Day 35 Day 56 Initial Day 7 Day 15 2% PVP 0 0.6 1.4 1.7 02.5 2.4 5% HP-β-CD 0 0.9 2.0 2.9 0 3.6 7.7 5% SBE-CD 0 2.5 5.5 7.1 0 7.412.5 0.1% CaCl₂ 0 0.4 0.9 1.5 0 2.2 2.4 CaCl₂/HPMC 0 0.6 1.4 1.9 0 2.55.5 0.5% HPMC 0 0.5 1.1 1.5 0 2.0 4.1 1% TRIS 0 0.7 1.6 1.7 0 3.0 7.8

Based on the stability data in the above Tables, it was concluded thatthe stability of compound I varies greatly over the pH range of 5-8.Further, increasing the solubility of compound I in SBE-cyclodextrinresulted in a decrease in the stability.

Example 5. Stability of 2.5%, 1.5%, 0.5% and 1.5% Ophthalmic Suspensionsof Compound I

The stability of a number of lots of compound I ophthalmic suspensions0.15%, 0.5%, 1.5% and 2.5% (initial pH of 7.34-7.7) as described belowin Table 18 were evaluated by monitoring the chemical, physical andmicrobiological stability characteristics of the product over time. Thechemical stability was evaluated by monitoring compound I and itsimpurities. The physical stability was evaluated by monitoring the pH,osmolality, viscosity, appearance, identity by X-ray Powder Diffraction(XRPD), and particle size. The microbiological stability was evaluatedby conducting sterility tests. A summary of the results (in ranges fromdifferent lots and various sampling times over the testing period) fromthe tests and stability monitoring limits for compound I ophthalmicsuspensions are presented in Table 13.

TABLE 13 Stability testing data for ophthalmic suspensions of compound ICondition monitored Total Impurity at Osmolality Compound I % impuritiesRRT of 1.2 pH Range: Storage suspension compound I (No more (No moreRange: 200-350 mOsm/kg condition (w/v) of label than 6.0%) than 4.5%)6.8-8.0 (initial 231-251) 25 ± 2° C./ 2.5% 98-100 0.0-0.2 <0.1-0.2 7.53-7.61 231-247 40% ± 5% 1.5% 99-100 0.1-0.2 0.1-0.2 7.52-7.63 243-244RH (up to 0.5% 96-98  0.2-0.5 0.2-0.5 7.49-7.51 242-243 26 weeks) 0.15% 95-99  0.3-1.6 0.3-1.6 7.54-7.60 236-244 40 ± 2° C./ 2.5% 98-103 0.2-0.60.2-0.6 7.52-7.58 235-240 <25% ± 1.5% 100-102  0.3-0.9 0.3-0.9 7.52-7.59244-255 5% RH 0.5% 97-97  0.7-2.4 0.7-2.4 7.48-7.52 246-251 (up to0.15%  85-95  1.2-8.6 1.2-8.6 7.45-7.56 239-252 26 weeks) 5 ± 2° C./2.5% 96-102 0.1-0.4 <0.1-0.2  7.45-7.59 230-245 35% ± 1.5% 98-1020.2-0.2 0.0-2.2 7.46-7.60 241-244 5% RH 0.5% 97-98  0.2-0.2 0.2-0.2 7.54240 (up to 0.15%  96-102 0.1-0.4 0.2-0.4 7.54-7.62 238-242 104 weeks)

In addition to the above parameters, viscosity was monitored and foundto be within 10% of the initial viscosity at each tested time pointunder each storage condition. Particle size measurements and sterilitywere also within stability monitoring limits. Further, the inventorsunexpectedly found that compound I that were micronized by ball millingin the final step of manufacturing maintained polymorphic form B withoutchanging crystalline character.

Example 6. Acute Toxicology Study of Exploratory Formulations

The four formulations from the exploratory stability studies in Table 2,FID 121522, FID 121511, FID 121512, FID 121513 were dosed in male NZWrabbits 5 times in one day, using a sterile irrigating solution as acontrol. No toxicity was observed for the four exploratory formulationsor the control.

Example 7. Pharmacokinetic Study of Exploratory Formulations

A pharmacokinetics study was conducted to determine the ocular uptake ofcompound I following single bilateral topical ocular dosing of male NZWrabbits with the four exploratory formulations provided in Table 2: FID121522, FID 121511, FID 121512, FID 121513. Results are shown in Table14.

TABLE 14 Summary of compound I C_(max) and C_(min) Results frompharmacokinetic study FID: 121522 121511 121512 121513 MatrixConcentration (nM) Cornea C_(max) 34700 15760 16400 901 C_(min) 39004520 1470 BLQ* Conjunctiva C_(max) 24500 4680 5270 165 C_(min) 1570 14202770 BLQ Aqueous Humor C_(max) 3730 1780 1940 185 C_(min) 98.9 55.4 20.1BLQ Iris-Ciliary Body C_(max) 3640 1480 1800 153 C_(min) 223 169 65 BLQPlasma C_(max) 63 30.2 43.5 0.834 C_(min) 2.18 1.00 0.741 BLQ IC₅₀ 30 3030 30 *BLQ = Below Limit of Quantitation Cmin determined at 4 hoursafter administration.

As seen in Table 14, the highest exposures of compound I was in thecornea, which was approximately 1.5 to 3 times the levels observed inthe conjunctiva, and approximately 10 times the levels observed in theaqueous humor and iris-ciliary body and approximately 500 times thelevels observed in plasma. In the cornea, C_(max) for the suspensionwith Poloxamer 407 was approximately 2 times the C_(max) for the twosuspensions with hypromellose and approximately 38 times the C_(max) forthe solution formulation. For the three suspensions, C_(max) in thecornea ranged from approximately 500 to 1200 times the IC₅₀ of 30 nM,while C_(min) ranged from approximately 50 to 130 times the IC₅₀.

Example 8. Pharmacokinetic Study of Further Formulations

Based on the results from the exploratory formulations, furtherformulations were prepared for additional pharmacokinetic studies on thecorneal and aqueous humor concentrations of compound I following singlebilateral topical ocular dosing in male pigmented rabbits. Theseformulations are shown in Table 15.

TABLE 15 Unpreserved Suspension Formulations of Compound I for PK StudyFID: 121746 121745 121744 Component Percent w/w Compound I 0.01 0.05 0.5Tyloxapol 0.05 0.05 0.05 Carbomer Homopolymer Type B 0.2 0.2 0.2Glycerin 2.0 2.0 2.0 Sodium Chloride 0.05 0.05 0.05 Tromethamine qs pH7.4 qs pH 7.4 qs pH 7.4 Hydrochloric Acid qs pH 7.4 qs pH 7.4 qs pH 7.4Purified Water qs 100 qs 100 qs 100

Results from the pharmacokinetic study are shown in Table 16.

TABLE 16 Summary of Cmax and Cmin Results from PK Study FID: 121746121745 121744 Matrix Concentration (nM) Cornea C_(max) 1150 5550 18500C_(min) 14 20 223 Aqueous Humor C_(max) 314 950 2990 C_(min) BLQ* BLQ11.1 IC₅₀ 30 30 30 *Below limit of quantitation Cmin results obtained at8 hours after dosing

As seen in Table 16, C_(max) and C_(min) for compound I show that thehighest exposures of compound I were in the cornea, with levelsapproximately 3 to 6 times the levels observed in the aqueous humor. Inthe cornea, C_(max) for the 0.5% suspension was approximately 3 timesthat of the 0.05% suspension and approximately 16 times that of the0.01% suspension. In the aqueous humor, C_(max) for the 0.5% suspensionwas approximately 3 times that of the 0.05% suspension and approximately9 times that of the 0.01% suspension. For the three suspensions, C_(max)in the cornea ranged from approximately 38 to 600 times the IC₅₀ of 30nM, while C_(min) ranged from approximately 0.5 to 7 times the IC₅₀.

A pharmacokinetics study was conducted to determine the corneal andaqueous humor concentrations of compound I following single bilateraltopical ocular dosing in intact eyes and following anterior keratectomywith and without bandage contact lenses in male pigmented rabbits usingthe formulations described in Table 15, FID121746, FID121745, andFID121744.

Results from the pharmacokinetic study are shown in Table 17.

TABLE 17 Summary of C_(max) and AUC_(0-8 h) Results from PK Study ofrabbits following anterior keratectomy FID: 121746 121745 121744 w/oCL^(a) w/o CL^(a) w/CL^(b) w/o CL^(a) w/CL^(b) Corneas Concentration(nM; AUC: nM · h) Intact C_(max) 1147 5550 18500 AUC_(0-8 h) 1433 455031700 Anterior C_(max) 1910 1490 10700 Keratectomy AUC_(0-8 h) 1770 440026700 IC₅₀ ^(c) 27 27 27 27 27 ^(a)Without bandage contact lenses^(b)With bandage contact lenses ^(c)IC50 for N-arachidonoyl dopamine(NADA)

Example 9. Toxicology Wound Healing Study with Compound I SuspensionFormulations

A toxicology wound healing study was conducted to evaluate corneal woundhealing following QID bilateral topical ocular dosing of compound Iafter unilateral laser photorefractive keratectomy in rabbits with andwithout bandage contact lenses. The compound I formulations used in thestudy are described in Table 18. For comparator purposes, ketorolactromethamine (ACULAR LS®) and dexamethasone (MAXIDEX®) formulations wereused.

TABLE 18 Compositions of Compound I Vehicle and Suspensions for WoundHealing Study FID: 121830 121744 121816 121814 Component Percent w/wCompound I 0.5 1.5 2.5 Tyloxapol 0.05 0.05 0.05 0.05 CarbomerHomopolymer 0.2 0.2 0.2 0.2 Type B Glycerin 2.0 2.0 2.0 2.0 Sodium 0.050.05 0.05 0.05 Chloride Tromethamine qs pH 7.4 qs pH 7.4 qs pH 7.4 qs pH7.4 Hydrochloric Acid qs pH 7.4 qs pH 7.4 qs pH 7.4 qs pH 7.4 PurifiedWater qs 100 qs 100 qs 100 qs 100

The study results demonstrated that after 3 days, the mean wound areasfor the corneas treated with 0.5, 1.5 and 2.5% compound I suspensionswith bandage contact lenses were smaller than those for the corneastreated with ACULAR LS and

MAXIDEX with bandage contact lenses, vehicle without bandage contactlenses and for untreated corneas. The mean wound areas for the corneastreated with 0.5, 1.5 and 2.5% compound I suspensions with bandagecontact lenses were comparable to those for the corneas treated withvehicle with bandage contact lenses and 2.5% compound I without bandagecontact lenses.

Example 10. Bacteriostasis/Stability Studies of Exemplary Compound ISuspensions

A series of five prototype sterile suspension formulations containing0.5% compound I were prepared and screened forbacteriostasis/fungistasis with five compendial organisms: S. aureus, P.aeruginosa, E. coli, C. albicans and A. brasiliensis. The criterion forstasis was no more than 0.5 log increase in microbial counts (CFU/mL).All the tested formulations demonstrated an acceptable level ofbacteriostasis/fungistasis. In addition, a 2.5% compound I suspension(otherwise identical to FID 121744), and suspension vehicle were alsoprepared and screened for bacteriostasis/fungistasis screening.

Bacteriostasis/fungistasis was observed through 3 days with an inoculumof ˜10⁶ and/or an inoculum of ˜10⁵.

Example 11. First in Human Study of Compound I

This example describes a first-in-human study of compound I conducted inhealthy volunteers. A total of 54 subjects were administered to studymedication. Part 1 of the first in human study administered singleascending doses of 0.15%, 1.5%, 2.5% w/v (1 drop) of compound I in aneye drop.

Part 2 tested multiple ascending doses (MAD) of compound I,administering (i) 1 drop of 0.15%, 1.5%, 2.5%, 4 times daily (every 6hours) for 7 days or (ii) 1 drop of 2.5%, 8 times daily (every 3 hours)for 7 days (supra-therapeutic dose) as eye drops.

Part 3 was an esthesiometry assessment to evaluate the anestheticproperties of compound I. Part 3 of the study had four arms, withcompound I, vehicle, tetracaine (0.5% ophthalmic solution, used as apositive control) and diclofenac sodium (0.1% ophthalmic solution, usedas the active NSAID comparator). Ocular anesthetic was selected becauseof its established anesthetic effect; however it is not the standard ofcare due to its negative effect on wound healing. NSAID is the currentstandard of care for corneal pain after PRK with minimal anesthetic butsignificant pain control properties. Vehicle eye drops were used as anegative control to enable better determination of the potentialanesthetic effect. Twelve healthy subjects, who met the eligibilitycriteria, were randomized to receive a single eye drop of 4 studytreatments, each during 1 of 4 different study days (Days 1, 4, 7 and10). Each subject was randomized to receive the study treatmentaccording to one of the following four sequences:

-   -   Sequence 1: Anesthetic, NSAID, compound I 2.5%, Vehicle    -   Sequence 2: NSAID, Vehicle, Anesthetic, compound I 2.5%    -   Sequence 3: Vehicle, compound I 2.5%, NSAID, Anesthetic    -   Sequence 4: compound I 2.5%, Anesthetic, Vehicle, NSAID

Safety assessments: Safety assessments consisted of collecting alladverse events (AEs), serious adverse events (SAEs), including systemicand ocular adverse events, along with their severity and relationship tostudy drug. Systemic safety assessments included regular monitoring ofhematology, blood chemistry and urinalysis performed at study center andregular assessments of physical examination, vital signs (systolic anddiastolic blood pressure, pulse rate, and body temperature), ECG,pregnancy and assessments of fertility and hand immersion test at 49° C.during MAD part. Ocular safety assessments included early treatmentdiabetic retinopathy study (ETDRS) visual acuity, intraocular pressure,slit-lamp biomicroscopy, corneal staining and dilated fundus exam.

Subjects were selected based on the following inclusion and exclusioncriteria.

Inclusion Criteria:

-   -   Written informed consent was obtained before any assessment was        performed.    -   Healthy male and female subjects aged 18 to 50 years        (inclusive), and in good health as determined by past medical        history, physical examination, vital signs, ECG, and laboratory        tests at Screening.    -   At Screening, and Baseline, vital signs (systolic and diastolic        blood pressure (BP) and pulse rate) were assessed in the sitting        position after the subject had rested for at least 3 minutes and        again after 3 minutes in the standing position. Sitting vital        signs were required to be within the normal range the following        ranges:    -   oral body temperature between 35.0-37.5° C.    -   systolic blood pressure (SBP), 90-150 mmHg    -   diastolic blood pressure (DBP), 50-90 mmHg    -   pulse rate, 40-100 bpm    -   Subjects were required to weigh at least 50 kg to participate in        the study, with a body mass index (BMI) within the range of        18-29 kg/m2. BMI=Body weight (kg)/[Height (m)]²    -   Subjects who were able to communicate well with the        Investigator, to understand and comply with the requirements of        the study.    -   For Part 3, subjects Baseline levels of eye sensitivity were to        be in the range of 50 to 60 mm (inclusive) as measured by the        Cochet-Bonnet esthesiometer.

Exclusion Criteria:

-   -   Women of child-bearing potential, defined as all women        physiologically capable of becoming pregnant, unless they were        using effective methods of contraception during dosing of study        treatment.    -   Subjects, who demonstrated any medical condition (systemic or        ophthalmic) that was, in the opinion of the Investigator, and        based on the content of the Investigator brochure, preclude the        safe administration of test article or safe participation in        this study.    -   Part 3 (esthesiometry): subjects who were using contact lenses        at the time of the study or had used in the past 3 years were        excluded to minimize variability in corneal sensitivity because        of contact lens use.    -   History of any ocular surgery or laser within the past 6 months        prior to Screening. History of any chronic eye disease other        than refractive error, incipient cataract, strabismic amblyopia,        or anisometropic amblyopia. Subjects with a history of acute eye        disease (such as infection, corneal abrasion or allergy) within        the past 6 months from Screening were eligible if the disease        was not active.    -   Any currently active ocular condition that required use of        topical eye drops.    -   Subjects using continuous positive airway pressure or other        sleep apnea devices.

Safety Results from the First in Human Study

Based on results from Parts 1, 2, and 3, the Maximum Tolerated Dose(MTD) was identified as the maximum feasible concentration of 2.5%, 8times daily for 7 days. No dose limiting adverse events were identifiedat this dose level. All adverse events of suspected causality tocompound I were of mild severity, except for moderate severity eyeirritation that lead to discontinuation of treatment in one patient ofthe 2.5% 4 times daily cohort. The most frequent ocular adverse eventsin the compound I treated patients were corneal staining, hyperemia andmild anterior chamber inflammation, in levels similar to placebo. Asummary of the adverse events from the SAD study are shown in Table 19.

TABLE 19 Overall incidence of AEs - number of events and number ofsubjects (Part 1: SAD) (Safety analysis set) Compound I Compound ICompound I 0.15% 1.5% 2.5% Vehicle Total N = 6 N = 6 N = 6 N = 6 N = 24nE, nS (%) nE, nS (%) nE, nS (%) nE, nS (%) nE,nS(%) AEs, Subjects withAEs  3, 1 (16.7)  3, 2 (33.3) 0, 0 (0.0) 4, 3 (50.0) 10, 6 (25.0) AEs ofMild severity  3, 1 (16.7)  3, 2 (33.3) 0, 0 (0.0) 3, 2 (33.3)  9, 5(20.8) AEs of Moderate severity 0, 0 (0.0) 0, 0 (0.0) 0, 0 (0.0) 1, 1(16.7) 1, 1 (4.2) Study drug-related AEs  3, 1 (16.7)  3, 2 (33.3) 0, 0(0.0) 3, 2 (33.3)  9, 5 (20.8) Serious AEs 0, 0 (0.0) 0, 0 (0.0) 0, 0(0.0) 0, 0 (0.0)  0, 0 (0.0) AEs leading to 0, 0 (0.0) 0, 0 (0.0) 0, 0(0.0) 1, 1 (16.7) 1, 1 (4.2) discontinuation of study Study-drug relatedAEs leading to 0, 0 (0.0) 0, 0 (0.0) 0, 0 (0.0) 1, 1 (16.7) 1, 1 (4.2)discontinuation of study N = number of subjects enrolled and receivedthe study drug nE = number of treatment emergent AE events in thecategory nS = number of subjects with at least one treatment emergent AEin the category Percent is based on the number of subjects

A summary of the adverse events from the SAD study are shown in Table20.

TABLE 20 Overall incidence of AEs - number of events and number ofsubjects (Part 2: MAD) (Safety analysis set) Compound I Compound ICompound I Compound I 0.15% 1.5% 2.5% 2.5% 4 times 4 times 4 times 8times daily daily daily daily Vehicle Total N = 6 N = 6 N = 6 N = 6 N =8 N = 32 nE, nS (%) nE, nS (%) nE, nS (%) nE, nS (%) nE, nS nE, nS (%)AEs, Subjects 6, 3 (50.0) 12, 5 (83.3) 4, 2 (33.3) 3, 2 (33.3) 13, 3(37.5) 38, 15 (46.9) with AEs AEs of Mild 6, 3 (50.0) 11, 5 (83.3) 3, 2(33.3) 3, 2 (33.3) 12, 3 (37.5) 35, 15 (46.9) severity AEs of Moderate0, 0 (0.0) 1, 1 (16.7) 1, 1 (16.7) 0, 0 (0.0) 1, 1 (12.5) 3, 3 (9.4)severity Study drug- 5, 2 (33.3) 9, 3 (50.0) 4, 2 (33.3) 2,1 (16.7) 10,3 (37.5) 30, 11 (34.4) related AEs Serious AEs (0.0) 0, 0 0, 0 (0.0) 0,0 (0.0) 0, 0 (0.0) 0, 0 (0.0) 0, 0 (0.0) AEs leading to 0, 0 (0.0) 0, 0(0.0) 1, 1 (16.7)^(a) 0, 0 (0.0) 0, 0 (0.0) 1, 1 (3.1) discontinuationof study Study-drug related 0, 0 (0.0) 0, 0 (0.0) 1, 1 (16.7) 0, 0 (0.0)0, 0 (0.0) 1, 1 (3.1) AEs N = number of subjects enrolled and receivedthe study drug nE = number of treatment emergent AE events in thecategory nS = number of subjects with at least one treatment emergent AEin the category Percent is based on the number of subjects ^(a)Subjecttreated with compound I 2.5% 4 times daily experienced eye discharge,ocular hyperemia (both of mild severity) and eye irritation (moderateseverity) in the left eye on Day 6., leading to discontinuation of theadministration of study drug.

For the hand immersion test, all subjects in the treatment cohortswithdrew their hand from water at 49° C. at a time interval between 0 to≤50 secs. No subjects lasted longer than 22 secs and no meaningfulchange in immersion time was observed between compound I andvehicle-treated subjects. The results therefore indicate that compound Idoes not alter temperature sensitivity of the subjects.

Esthesiometry Study (Part 3)

The first-in-human study further evaluated potential anesthetic effectof topical ocular 2.5% compound I by esthesiometry testing, i.e.measurement of the filament length (cm) when the threshold of corneatouch is perceived.

An analysis of the results showed that tetracaine 0.5% had an anestheticeffect with approximately 10 minutes of duration of action (positivecontrol). Both diclofenac 0.1% and vehicle had no anesthetic effect onthe cornea, as expected based on known clinical experience withdiclofenac. Compound I 2.50% showed no anesthetic effect at any timepoint after treatment.

For measures of corneal sensitivity, statistical and clinicalsignificance were observed while comparing least squares means (Test vsRef) for the following:

-   -   Tetracaine 0.5% (N=12) vs Vehicle (N=12) at 2.5 minutes, 10        minutes and 20 minutes post-dose;    -   Tetracaine 0.5% (N=12) vs Diclofenac 0.1% (N=12) at 2.5 minutes,        10 minutes and 20 minutes post-dose;    -   Compound I 2.5% (N=11) vs Tetracaine 0.5% (N=12) at 2.5 minutes        and 10 minutes post-dose

No difference was seen in corneal sensitivity between compound I 2.5%and vehicle, with all p-values greater than or equal to 0.395 atscheduled time points up to 30 minutes post-dose. This studydemonstrated a lack of anesthetic effect of 2.5% compound I whencompared to tetracaine 0.5% (drug with anesthetic effect as positivecontrol) and vehicle (placebo).

Pharmacokinetic Summary from First in Human Study

After both single and multiple topical ocular suspension doses ofcompound I 0.15%, 1.5%, and 2.5%, plasma PK profile showed rapidabsorption of compound I into systemic circulation and low concentrationexposures achieved with moderate variability across subjects. Increasein dose from 0.15% to 2.5% resulted in less than dose-proportionalincrease in systemic exposure. Accumulation of compound I atsteady-state was minor (˜1.7-fold) following administration of 0.15%,1.5% or 2.5% suspension 4 times daily and also minor (˜1.3-fold)following 2.5% 8 times daily for 7 days.

Example 12. Clinical Study of Compound I for Treatment of PostoperativePain

This example describes a clinical study of compound I(4-(7-hydroxy-2-isopropyl oxo-4H-quinazolin-3-yl)-benzonitrile) in thetreatment of postoperative ocular pain in patients undergoingphotorefractive keratectomy (PRK) surgery. The formulation used isillustrated in Table 18.

Study Objectives

Primary objective Endpoints related to primary objective To evaluatepain control in the Visual analog scale (VAS) pre-dose pain assessmentimmediate post-operative period. at 6 hours post-operatively Averageocular pain VAS assessments from the first post-operative assessment upto the pre-dose 12 hour assessment Secondary objectives Endpointsrelated to secondary objectives To evaluate the efficacy of compoundIncidence and amount of rescue oral analgesics I 2.5% eye drops fourtimes daily for needed in 6 hours, 12 hours, 24 hours, 2 days, and 3reducing use of oral analgesics days post-operatively after each PRKsurgery. following PRK procedure To assess safety and tolerability ofAdverse events (AEs) and serious adverse events compound I 2.5% eyedrops four (SAEs) times daily Visual acuity, intraocular pressure (IOP),dilated fundus exam, ocular hyperemia Size of epithelial defect by slitlamp exam Blink rate, tear production Vital signs (blood pressure, pulserate, and body temperature) To evaluate pain severity post- All VASmeasurements during the first 3 days after surgery operatively To assessthe systemic exposure Plasma concentration of compound I after oculardosing of compound I 2.5% eye drops four times daily at various timepoints in PRK patients Exploratory objectives Endpoints related toexploratory objectives To explore wound healing rate Size of epithelialdefect by anterior segment Optical coherence tomography (OCT) To exploreeffect on pain relief of VAS scores before and after instillation of eyedrop compound I versus Vehicle after (three time points) instillation ofeye drop To explore effect of compound I on Ocular Pain AssessmentSurvey (OPAS) ocular pain and quality of life compound I exposure in thebandage Residual compound I amount after treatment contact lens (BCL)

Study Design

The study was a proof of concept, double-masked, randomized,vehicle-controlled study of compound I administered as eye drops inaddition to standard of care treatment in patients after PRK surgery.PRK surgery was performed as an outpatient procedure under topicalanesthesia with removal of the corneal epithelium to expose the stromafor the laser ablation. Compound I dosing was as single eye drop of 2.5%(0.925 mg/drop), administered four times daily (every six hours) in oneeye from immediate post-op (time 0) to last dose at 72 hours.

Vehicle dosing was as a single drop, administered four times daily(every six hours) in one eye from immediate post-operative (time 0) tolast dose at 72 hours.

The study consisted of 2 treatment periods using a crossover design.Patients underwent PRK surgery on 2 separate occasions (periods), oneeye at a time. Patients were randomized to receive either compound 1 orvehicle following procedure 1 and the alternate following procedure 2.

Forty patients were randomized in a 1:1 ratio to two sequences: compoundI during period 1, followed by vehicle in period 2, or vehicle duringperiod 1 followed by compound I during period 2. Each patient wasadministered one drop four times daily for 72 hours following PRKsurgery on their study eye (Day 1, Periods 1 and 2). The initial studyeye was the non-dominant eye as established at screening and inagreement of the patient and the investigator. The patients returned forfollow-up visits on Days 2, 3, 4 and 8 of Period 1 after surgery in thefirst eye, with optional daily visits to follow the patient until woundhealing was complete. The second eye surgery was not performed if anycomplications were noticed.

The patients underwent PRK surgery on their second study eye (dominanteye) on Period 2, Day 1. The PRK surgery was performed after theepithelial defect of the first eye was resolved and at the discretion ofthe Investigator. Following the PRK surgery, the patients received theopposing treatment four times daily for 72 hours. Patients returneddaily for the first 3 postoperative days (Period 2, Days 2-4) and at oneweek after the second surgery (Period 2, Day 8), with optional dailyvisits to follow the patient until wound healing was complete. An end ofstudy (EOS) visit took place 30 days after the second eye surgery (orafter final dose of investigational product when the patient endedtreatment early in Period 1).

All patients received standard of care treatment during periods 1 and 2of the study, including application of a bandage contact lens (AirOptix® Night and Day® Aqua or equivalent) following the PRK procedureand before receiving study drops for pain. A course of topical ocularantibiotic (Moxifloxacin or equivalent 1 eye drop four times daily) wasstarted after application of first dose of study drops and was continuedfor 4-7 days. Prednisolone acetate ophthalmic one eye drop four timesdaily was administered for one week after PRK, followed by taper.Preservative-free unit-dose artificial tears were used as needed. Thefirst dose of study drops after each PRK procedure was administered bysite staff. Subsequent doses were self-administered. A gap of 5 minuteswas allowed between eye drop administrations in sequence. To summarize,patients underwent PRK procedure, bandage lens was placed on the cornea,compound I or Vehicle was administered; after approximately 5 minutesantibiotic was administered, and after another 5 minutes, prednisolonewas administered.

Rescue medication consisted of oral analgesic (acetaminophen 300mg+codeine 30 mg) as needed up to a total of 10 tabs/day or 1-2 tabsevery 4 hours.

Additional Study Treatment

All patients received standard of care ancillary treatment following PRKsurgery, including:

-   -   Bandage contact lens (Air Optix® Night and Day® Aqua or        equivalent)    -   A course of topical ocular antibiotic (Moxifloxacin or        equivalent 1 eye drop four times daily) was started right after        PRK surgery, and after application of first dose of study eye        drop and continued for 4-7 days, per managing physician.    -   Prednisolone acetate ophthalmic: one eye drop four times daily        was started immediately following PRK surgery and after        instillation of antibiotic eye drop and was administered for 1        week after PRK, followed by taper per local procedures.    -   Preservative-free unit-dose artificial tears were used as        needed. Artificial tears were not allowed to be chilled for        analgesic effect.

The drops were administered in sequence, with a gap of at least 5minutes between eye drop administrations.

Inclusion Criteria

Population eligible for inclusion in this study had to fulfill all ofthe following criteria:

-   -   Male and female patients aged 18 to 75 years eligible for        bilateral PRK surgery.    -   Normal eye exam except for refractive error at Baseline.    -   Planned myopia correction was required not to exceed −4.00        Diopters (sphere) and 3.00 diopters of astigmatism, with        spherical equivalent not higher than ˜4.50, confirmed by        manifest refraction at Baseline. Monovision treatment (such as        correction for far distance in one eye and for intermediate        distance in the fellow eye) was allowed.    -   Written informed consent was obtained before any assessment was        performed.

Exclusion Criteria

-   -   Monocular patient (including amblyopia) or best corrected visual        acuity (BCVA) score worse than 20/80 (Snellen) or 55 letters        [early treatment diabetic retinopathy study (EDTRS)] at        Baseline.    -   Any systemic or ocular disease that affected wound healing (such        as severe rheumatoid arthritis or diabetes or history of keloid        formation) or a history of ocular trauma, uveitis, infection, or        inflammation in the 6 months prior to Baseline. Especially for        diabetes: Patients with severe diabetes, uncontrolled diabetes,        diabetic keratopathy, diabetic retinopathy, diabetic macular        edema, diabetic nephropathy, diabetic foot ulcers or other        systemic complications of diabetes were excluded. Patients with        mild, well-controlled diabetes with no evidence of ocular or        systemic complications of diabetes were included.    -   Patients with active inflammatory or infectious ocular        conditions, severe or progressive retinal disease, and use of        topical or systemic steroids, or use of Coumadin or similar        drugs within the last 6 months prior to Baseline.    -   Patients with any corneal dystrophy (epithelial, stromal or        endothelial) or any cornea disease (including significant        scarring (at the discretion of the Investigator), ocular herpes        or pterygium).    -   Previous refractive or corneal surgery (such as LASIK, PRK,        radial keratotomy, pterygium removal, corneal transplantation).    -   History of allergic or hypersensitivity reaction or significant        AEs to any of the drugs used in this study including tetracaine        or similar topical ocular anesthetic, NSAIDs and aspirin, oral        analgesic (including acetaminophen and codeine), antibiotics,        steroids and inability to tolerate or wear bandage contact lens    -   Concurrent therapy or history of chronic therapy or abuse of        systemic or ocular NSAIDs, analgesics, pain medication        (including gabapentin or pregabalin and similar), opiates or        cannabis.    -   Patients who used any topical eye medication except for        lubricating eye drops within two weeks prior to surgery in the        study eye were excluded. Patients meeting any of the following        were excluded:        -   Usage of topical NSAIDs during 30 days before Baseline, OR        -   Systematic/chronic usage of systemic NSAIDS within 30 days            prior to Baseline, OR        -   Occasional usage of systemic NSAIDS within 3 days prior to            Baseline, OR        -   Usage of ocular cyclosporine (or similar medication) within            the 3 months prior to surgery.    -   Patients with body weight <50 kg, or who do not have a body mass        index (BMI) within the range of 18-35 kg/m². BMI=Body weight        (kg)/[Height (m)]²    -   Pregnant or nursing (lactating) women. Women of child-bearing        potential, defined as all women physiologically capable of        becoming pregnant, unless they are using basic methods of        contraception during dosing of investigational drug.

No additional exclusions were applied by the Investigator, in order thatthe study population was the representative of all eligible patients.

Study Population

The study population was comprised of male and female patients eligiblefor PRK surgery between 18 and 75 years old (inclusive). A total of 40patients were planned. A total of 44 patients were screened and of them40 patients were enrolled in the study and randomized.

Patient selection was established based on a review of all eligibilitycriteria at Screening and Baseline. A relevant record (e.g. checklist)of the eligibility criteria was stored with the source documentation atthe study site. Deviation from any entry criterion excluded a patientfrom enrollment into the study.

Patient demographics are provided in Table 21.

TABLE 21 Patient demographics by treatment sequence (Safety analysisset) Vehicle/ compound I compound I 2.5%/ 2.5% Vehicle Total N = 20 N =20 N = 40 Age (years) Mean (SD) 34.4 (10.77)  33.7 (8.94)  34.0 (9.78) Median 33.5 32.0 33.0 Range 20-54 23-56 20-56 Sex - n (%) Male 11 (55%)10 (50%) 21 (53%) Female  9 (45%) 10 (50%)  19(48%) Race - n (%) White18 (90%) 17 (85%) 35 (88%) Asian  2 (10%) 1 (5%) 3 (8%) Black Or 0  1(5%) 1 (3%) African American Multiple 0  1 (5%) 1 (3%) Ethnicity - n (%)Not Hispanic 19 (95%) 18 (90%) 37 (93%) Or Latino Hispanic Or 1 (5%)  2(10%) 3 (8%) Latino Weight (kg) Mean (SD) 84.1 (14.86)  78.8 (14.07) 81.5 (14.53)  Median 84.7 76.1 79.2 Range  60-119  57-107  57-119 Height(cm) Mean (SD) 170.4 (7.40)    173.2 (6.96)    171.8 (7.23)    Median171.3  174.1  172.7  Range 155-184 161-185 155-185 BMI (kg/m²) Mean (SD)28.9 (3.98)  26.2 (3.73)  27.5 (4.04)  Median 29.7 26.0 27.5 Range 21-3521-35 21-35

Treatment Arms

Patients were assigned to one of the following 2 treatment sequences ina ratio of 1:1 lasting 3 days per period.

Sequence Period 1 Period 2 1 Compound I 2.5% Vehicle control 4 timesdaily (every 6 hours) 4 times daily (every 6 hours) for 72 hours(inclusive) for 72 hours (inclusive) 2 Vehicle control Compound I 2.5% 4times daily (every 6 hours) 4 times daily (every 6 hours) for 72 hours(inclusive) for 72 hours (inclusive)

Compound I was administered to the patient as ocular drops. Drops wereadministered at the study site by the study personnel during the day ofPRK surgery. During the daily follow-up postoperative visits, when thepatient was at the site during the expected time for eye dropadministration, the study personnel was required to administer the eyedrops. The remainder of drops were home administered by the patient, orwhen the patient was present at the study site for a visit (e.g., at 24,48 and 72 hours post-surgery).

Visual Acuity Scale

Patient subjective experience of pain was recorded using VAS, a numericassessment of pain between 0 and 100, with 0 representing no pain, and100 representing worst imaginable pain. Previous studies of pain in PRKshowed that the most intense pain was experienced within the first 12hours after surgery with the peak around 4-6 hours after surgery (Sheret al., Refract Corneal Surg. November-December; 9(6):425-36 (1993)),and these were the time points selected for primary endpoint analysis.Since it was important clinically to both decrease the maximum pain aswell as the overall pain that the patient experiences during theimmediate postoperative period, the two periods of 6 hours and up to 12hours postoperatively, were evaluated as primary endpoints. Allevaluable VAS data was collected using an ePRO, which was an electronicdevice (cell phone with software application), on which patients markedtheir pain levels at the appropriate time points.

Rescue oral analgesics: It is not ethical to refuse pain controlmedication to patients postoperatively as part of a clinical trial. Areview of prior clinical trials indicated that NSAIDs are used forpostoperative pain after PRK as rescue oral analgesics, similar to theStandard of care after PRK. Because the use of oral rescue medicationmay be a potential confounder of the pain VAS evaluation, threeapproaches were used when analyzing pain VAS scores to account for theinfluence of pain meds (assuming 4 hours of rescue medication effect):(1) any recorded VAS score within 4 hours after use of rescue medicationwas considered missing; (2) all the recorded VAS scores was used; and(3) any recorded VAS scores within 4 hours after rescue medication usewas imputed by the record taken prior to the rescue medication.

Primary Efficacy Results

The mean VAS pain severity scores at 6 hours and 0-12 hourspost-operatively are presented in Table 22 and Table 23, respectively,and shown in FIG. 3 . The primary analysis was performed using thePrimary PD analysis set.

The treatment differences in mean VAS pain severity scores at 6 hoursand over 0-12 hours period post-operatively between compound I andVehicle were statistically significant with p-values less than 0.10.

The model based means treatment difference (compound I—Vehicle) in VASpain severity scores at 6 hours post-operatively was −11.1 (90% CI:(−17.54, −4.71; p=0.005) and at 0-12 hours period post-operatively was−8.56 (90% CI: (−14.29, −2.83; p=0.016). Thus, the primary efficacyobjective of the study was met.

The treatment differences in mean VAS pain severity scores at 6 hoursand over 0-12 hours period post-operatively between compound I andVehicle were statistically significant with p-values less than 0.10.

TABLE 22 Mean VAS pain severity scores at 6-hours post-operatively(Primary PD analysis set) Model estimated mean* Comparison of modelbased means: (SE) compound I vs. Vehicle Diff compound I Vehicle(compound I − (N = 30)^(#) (N = 29) Vehicle) % Diff 90% CI P-value 34.63(4.05) 45.76 (4.10) −11.1 −25% (−17.54, 0.005 −4.71) *To account fororal rescue medication (ORM) use, any recorded VAS pain scores taken upto 4 hours after the use of ORM was replaced with the VAS score recordedjust before use of ORM, per FDA guidance 2014. ^(#) ePRO data from thefirst 10 patients (out 40 total) were not evaluable due to failure ofthe first vendor/ePRO device

TABLE 23 Mean VAS pain severity scores 0-12 hours post-operatively(Primary PD analysis set) Model estimated mean* Comparison of modelbased means: (90% CI) compound I vs. Vehicle Diff compound I Vehicle(compound I − (N = 30) (N = 29) Vehicle) % Diff 90% CI P-value 30.9039.46 −8.56 −22% (−14.29, 0.016 (24.04, (32.54, −2.83) 37.76) 46.39) *Toaccount for oral rescue medication (ORM) use, any recorded VAS painscores taken up to 4 hours after the use of ORM was replaced with theVAS score recorded just before use of ORM, per FDA guidance 2014.

Secondary Efficacy Results

The number of patients who did not use oral rescue medication (ORM) washigher in compound I-treated eye compared to the Vehicle-treated eye at0-6 hours, 0-12 hours and 0-24 hours post-operatively. After 36 hourspost-operation, the same number of patients took ORM in compound I vs.Vehicle (Table 24).

TABLE 24 Summary of oral rescue medication use incidence (number ofpatients who did not use oral rescue medication) (Secondary PD analysisset) compound I 2.5% Vehicle N = 40 N = 40 Time interval n (%) n (%)p-value 0-6 hours post-operatively 23 (57.5%) 19 (47.5%) 0.2891 0-12hours post-operatively 19 (47.5%) 16 (40.0%) 0.5811 0-24 hourspost-operatively 16 (40.0%) 12 (30.0%) 0.3438 0-2 days post-operatively11 (27.5%) 11 (27.5%) 1.0000 0-3 days post-operatively 11 (27.5%) 11(27.5%) 1.0000 n: Number of patients who did not use oral rescuemedication (ORM) * For the both treatment sequences column, n representsthe number of patients who did not use oral rescue medication (ORM) inany treatment group.

Histogram of oral rescue medication (ORM) use incidence (number ofpatients who did not use ORM) is displayed in FIG. 4 .

The summary and analysis of amount of ORM (number of pills per patient)is presented in Table 25.

TABLE 25 Summary of amount of oral rescue medication (Number of pills)(Secondary PD analysis set) % Change compound I Vehicle (Compound I −Vehicle)/ Time interval Statistics N = 40 N = 40 Vehicle 0-6 hours Mean(SD) 0.75 (1.032) 1.00 (1.132) −25% post-operatively Range 0.0-4.0 0.0-4.0  p-value 0.10* 0-12 hours Mean (SD) 1.40 (1.780) 1.65 (1.847)−15% post-operatively Range 0.0-6.0  0.0-6.0  p-value 0.26  0-24 hoursMean (SD) 2.35 (2.751) 2.80 (3.006) −16% post-operatively Range 0.0-10.00.0-12.0 p-value 0.05* 0-48 hours Mean (SD) 4.05 (4.466) 4.68 (5.225)−13% post-operatively Range 0.0-16.0 0.0-20.0 p-value 0.05* 0-72 hoursMean (SD) 4.33 (4.896) 5.05 (5.574) −14% post-operatively Range 0.0-21.00.0-22.0 p-value 0.07* *Obtained from a Wilcoxon signed rank test N =total number of patients at each treatment *Statistical significancewith p ≤ 0.10 (per primary endpoint power calculation)

As seen in Table 25 and FIG. 4 , in every time interval during the studythere was less ORM taken during compound I vs. Vehicle treatment

The summary and analysis of amount of ORM (mg/kg of body weight) ispresented in Table 26.

In every time interval, amount of ORM (mg/kg of body weight) was lessduring compound I vs. Vehicle treatment (Table 26). During the periods0-6 hours, 0-24 hours, 0-48 hours and 0-72 hours post-operatively thedifference in milligrams per kilogram of body weight was statisticallysignificant (p≤0.10).

TABLE 26 Summary of amount of oral rescue medication (mg/kg of bodyweight) (Secondary PD analysis set) % Change Time Compound I Vehicle(Compound I − Vehicle)/ interval Statistics N = 40 N = 40 Vehicle 0-6hours Mean (SD) 3.32 (4.684) 4.41 (5.038) −25% post-operatively Range0.0-19.2 0.0-17.1 p-value 0.09* 0-12 hours Mean (SD) 6.10 (8.062) 7.39(8.478) −17% post-operatively Range 0.0-28.7 0.0-28.7 p-value 0.21  0-24hours Mean (SD) 10.17 (12.335) 12.41 (14.130) −18% post-operativelyRange 0.0-47.9 0.0-57.5 p-value 0.05* 0-48 hours Mean (SD) 17.59(19.850) 20.51 (23.991) −14% post-operatively Range 0.0-76.6 0.0-95.8p-value 0.05* 0-72 hours Mean (SD) 18.70 (21.509) 22.06 (25.527) −15%post-operatively Range 0.0-87.4  0.0-105.4 p-value 0.06* *Obtained froma Wilcoxon signed rank test N = total number of patients at eachtreatment *Statistical significance with p ≤ 0.10 (per primary endpointpower calculation) Each acetaminophen/codeine tablet is considered as330 mg in the calculation of amount (mg/kg of body weight).

VAS Pain Severity During the First 3 Days after PRK Surgery

After PRK surgery, patients reported statistically significantly lowerVAS pain severity scores after treatment with compound I compared toVehicle at p-value threshold 0.10 in 6 out of the 7 time points checkedduring the first 18 hours after surgery, starting with the hour 1 aftersurgery. The VAS pain severity scores were lower for compound I thanVehicle at all scheduled time points up to and including 36 hours aftersurgery. At all-time points from 36 hours until the end of the VAScollection period at 72 hours after PRK surgery, the difference betweencompound I VAS scores and Vehicle VAS scores was only marginallydifferent, and there was no statistically significant difference betweenthe scores at the p-value threshold of 0.10.

VAS Pain Severity Scores Before and After Instillation of Study Eye Drop

The mean changes in VAS scores from time points 6.5, 18.5 and 24.5 hourspost-operative compared one-half hour prior to that and immediatelybefore instillation of eye drops (namely at hours 6, 18 and 24) were−3.1, 2.8 and 1.3, respectively for compound I-treated eyes. ForVehicle-treated eyes the same differences were −5.6, 2.2 and −0.2,respectively.

Ocular Pain Assessment Survey (OPAS)

The OPAS is a validated instrument for quantifying and monitoringcorneal and ocular surface pain and quality of life, developed inresponse to an identified need from a National Eye Institute Workshop in2010. See Qazi et al., Ophthalmology July 123(7):1458-1468 (2016). Therating scale of the overall pain severity in the OPAS from Qazi et al.rated from 0 (no pain) to 10 (severe pain) or for frequency of symptomsfrom 0% (never) to 100% (all the time), as per the survey. The patientswere asked to fill out the OPAS survey/questionnaire at Day 2, Day 4 (atthe end of treatment period with study drug) and Day 8. Statisticalanalysis of OPAS results was not performed. Of the total of 27 OPASquestions, the results of seven questions are presented below.

In the questions of eye pain intensity during the previous 24 hours atthe level of eye pain that is the most painful (question 4), level ofpain that is the least painful (question 5) and level of eye pain inaverage (question 6) at Day 2 all 3 answers numerically favored compoundI compared to Vehicle (FIG. 5A, FIG. 5B, FIG. 5C).

In the questions of how often eye pain was associated by the followingsymptoms: redness (question 22), burning (question 23), sensitivity tolight (question 24) and tearing (question 25), all answers favoredcompound I compared to Vehicle (FIGS. 6A, 6B, 6C, and 4D). Thus,patients administered compound I exhibited lower levels of eye painassociated with redness, burning, sensitivity to light (photophobia),and tearing compared to patients administered placebo.

Summary Exploratory Ocular Results

-   -   Ocular Pain Assessment Survey (OPAS) showed better pain control        and quality of life patients during the compound I vs. the        Vehicle treatment period.    -   In the questions of eye pain intensity during the previous 24        hours at the level of eye pain that is the most painful        (question 4), level of pain that is the least painful        (question 5) and level of eye pain in average (question 6) at        day 2 all 3 answers favored compound I compared to Vehicle        (FIGS. 5A-5C).    -   In the questions of how often eye pain was associated by the        following symptoms: redness (question 22), burning (question        23), sensitivity to light (question 24) and tearing (question        25), all answers favored compound I compared to Vehicle (FIGS.        6A-6D).    -   The VAS scores from time points 6.5, 18.5 and 24.5 hours        post-operatively compared one-half hour prior to that and        immediately before instillation of eye drops (namely at hours 6,        18 and 24) were −3.1, 2.8 and 1.3, respectively for compound        I-treated eyes. For Vehicle-treated eyes the same differences        were −5.6, 2.2 and −0.2, respectively.

Pharmacokinetics

Pharmacokinetic Assessments

Pharmacokinetic (PK) samples were collected at the time points definedin the visit schedule supra, with PK blood collection windows shown inTable 27.

TABLE 27 Permitted time window for PK blood collection Pre-dosePost-dose Within 90 min prior to dosing Within ±5 min of expected timepoint (sample numbers 101. 106, 107, relative to dose, up to 2 hourspost-dose 112, 117 and 118) Within ±5 min from 72.25 to 74 hours (0.25to 2 hours relative to timing of last dose)

All blood samples (3 mL) were taken from the arm by either directvenipuncture or an indwelling catheter inserted in a forearm vein. Aftereach tube of blood was drawn, it was immediately inverted gently 8-10times to ensure the mixing of tube contents with anticoagulant (3 mL K2EDTA). Prolonged sample contact with the rubber stopper was avoided andthe tubes were placed upright in a test tube rack surrounded by wet iceuntil centrifugation.

Within 30 minutes, the sample was centrifuged at about 5° C. for 10minutes at approximately 2000 G (or the sample was placed on ice andcentrifuged at room temperature). Immediately after centrifugation, thewhole supernatant (approx. 1.5 mL) was transferred in the first 1.8 mLNUNC 2D barcoded cryovial. After mixing the plasma thoroughly, half ofthe plasma was transferred from the first cryovial to the secondcryovial and the caps were secured. Appropriate PK cryolabels wereattached to each cryovial and the labels were secured with clear tape.The cryovials were frozen immediately over dry ice then, kept frozen at≤−20° C. until shipment to the central lab. The vials were shipped inbiweekly batches.

Compound I was quantified in plasma using a validated LC-MS/MS method;the lower limit of quantification (LLOQ) was 0.05 ng/mL. Concentrationswere expressed in ng per mL units. When feasible, bandage contact lenses(BCL) exposed to study drug were collected and analyzed for residualdrug exposure after treatment (LLOQ: 5.00 ng/mL in 0.55 mL of extractionfluid, or 2.75 ng/BCL). Concentrations below the LLOQ were reported as“zero” and missing data were labeled as such.

The following PK parameters were determined as relevant using the actualrecorded sampling times and non-compartmental method with Phoenix™WinNonlin® (Version 6.4): Cmax, Tmax, AUClast (calculated), Clast andTlast from the plasma concentration-time data. Pre-dose concentrationswere determined by inspection at nominal 0, 24 and 72 hours. The lineartrapezoidal rule was used for AUClast calculation. No PK parameters werecalculated from the bandage contact lenses compound I concentrationdata.

Plasma Pharmacokinetics of Compound I

Arithmetic mean plasma concentration-time profiles and PK parameters ofcompound I are presented in FIG. 7A and FIG. 7B and Table 28.

After the unilateral topical ocular dose of compound I, absorption intothe systemic circulation was rapid, with median Tmax of 0.459 hr and0.467 hr after the first (Day 1; range 0.167-2.00 hr, FIG. 7A) and 13th(Day 4; range 0.00-2.08, FIG. 7B) doses, respectively. Cmax wasdetermined and AUClast calculated for 34/40 patients on Day 1 and 33/40(Cmax) or 31/40 (AUClast) patients on Day 4. All Day 1 pre-doseconcentrations were below the limit of quantitation and imputed as 0.00ng/mL. Trough (end of 6-hour dose interval and prior to next dose) meanconcentration (CV %) prior to the 5th dose on Day 2 (24 hr post 1stdose; Day 2 time point of 0 h) was 1.25 ng/mL (54.9%), and slightlyhigher at 1.99 ng/mL (76.3%) prior to the 13th dose on Day 4 (72 hr post1st dose; Day 4 time point of 0 h).

Concentrations were generally low, with observed Cmax ranging from 0.195to 7.56 ng/mL, and AUClast from 0.261 to 14.5 ng*hr/mL, across 4 days ofrepeated QID dosing. Arithmetic mean Cmax (CV %) following the firstdose on Day 1 was 0.454 ng/mL (49.9%) and 2.40 ng/mL (63.5%), i.e.5.3-fold higher, after the 13th dose on Day 4. Corresponding meanAUClast (CV %) values were 0.638 ng*hr/mL (46.0%) and 4.38 ng*hr/mL(67.0%), representing a 6.9-fold increase over the 4 days.

TABLE 28 Summary statistics of compound I PK parameters after topicalocular administration of 2.5% compound I (PK analysis set) Profile CmaxTmax AUClast day Statistic (ng/mL) (hr) (ng*hr/mL)¹ 1 N 34 34 34 Mean(SD) 0.454 (0.227) — 0.638 (0.293) CV % 49.9 — 46.0 Median 0.375 0.4590.569 Min, Max 0.195, 1.05 0.167, 2.00 0.261, 1.58 4 N 33 33 31 Mean(SD) 2.40 (1.53) — 4.38 (2.94) CV % 63.5 — 67.0 Median 2.07 0.467 3.42Min, Max 0.639, 7.56  0.00, 2.08  1.18, 14.5 ¹Median Tlast (range): Day1, 2.00 hr (1.95-2.05 hr); Day 4, 2.00 hr (1.77-2.10 hr) Clast is notshown.

Bandage Contact Lens

Bandage Contact Lens (BCL) was collected from 40 patients on Day 4 ofthe dose administration and analyzed for compound I. Values from thethree IMP non-compliant patients were excluded from BCL summarystatistics, leaving 37 values evaluable, including one value from apatient without available plasma PK. The mean concentration of compoundI (CV %) in 0.55 mL extraction fluid was 8500 ng/mL (73.1%), with a widerange of 284 to 22600 ng/mL. These concentrations translate to anestimated mean of 4680 ng/lens (0.55 mL×8500 ng/mL), and a range of 156to 12400 ng/lens. The amount in BCL was very small in relation to the0.925 mg dose of compound I administered: mean 0.51%, and range of0.017% to 1.3% of dose. This indicates that the BCL absorbs and retainstopical ocular compound I, but at negligible amounts compared to thetopical ocular dose.

Summary Pharmacokinetic Results

-   -   Absorption of compound I into the systemic circulation was rapid        (median plasma Tmax was approximately 0.5 hours) after either        single (first dose) or repeated 4 times daily topical ocular        administration.    -   After both single and multiple topical ocular dose        administration of compound I, the systemic exposure was low,        ranging from 0.195 ng/mL to 7.56 ng/mL across 4 days of repeated        QID dosing, and showed moderate variability. Mean Cmax was 0.454        ng/mL after the first dose on Day 1 and 2.40 ng/mL after the        13^(th) dose on Day 4.    -   The CV % values for Cmax and AUClast following single dose were        49.9% and 46.0%, respectively. The corresponding values after        repeated administration were 63.5% and 67.0%.    -   Mean Cmax after repeated administration (2.40 ng/mL) was        5.3-fold higher than after single dose on Day 1 (0.454 ng/mL).        Similarly, mean AUC last was 6.9-fold higher after repeated        versus single administration (4.38 versus 0.638 ng*hr/mL). These        data indicated accumulation over the 4 day period of 2.5%        compound I administration 4 times daily.    -   Bandage Contact Lens (BCL) was collected from 40 patients on Day        4 of dose administration. The mean concentration (CV %) in 0.55        mL extraction fluid was 8500 ng/mL (73.1%), which converts to an        estimated 4680 ng/lens. The amount in BCL was very small in        relation to the 0.925 mg dose of compound I administered: 0.51%,        and range of 0.017% to 1.3% of dose. This indicates that the BCL        absorbs and retains topical ocular compound I, but at negligible        amounts compared to the topical ocular dose.

Safety

Safety assessments consisted of collecting all AEs, SAES, with theirseverity and relationship to study drug. Table 29 includes a list ofsigns and symptoms which are common in the post-PRK surgery setting.Only signs and symptoms greater than the listed ranges of eitherseverity or duration were reported as AEs.

TABLE 29 Signs and symptoms of PRK surgery which do not requirereporting Expected Expected duration Sign/symptom severity after PRKsurgery Corneal epithelial defect Mild to moderate 1 week Eve Edema Mildto moderate 1 week Eyelid Edema Mild to moderate 1 week PunctateKeratitis Mild to moderate 1 week Conjunctival Hyperemia Mild tomoderate 1 week Reduction in BCVA of 10 letters Mild to moderate 1 monthor more from baseline Eye pain Mild to moderate 1 week Ocularirritation/discomfort Mild to moderate 1 month Foreign body sensationMild to moderate 1 month Lid margin irritation/crusting Mild to moderate1 month Increased Lacrimation Mild to moderate 1 month Vision that isshadowy Mild to moderate 1 month Vision that is hazy Mild to moderate 1month Blurred vision Mild to moderate 1 month Photophobia Mild tomoderate 1 month Dry eye Mild to moderate 1 month Glare Mild to moderate1 month Halos Mild to moderate 1 month

One week indicates resolution by the Day 8 post-op visit (Visit 6 intreatment period 1 or Visit 11 in period 2), including the allowablewindow. The table was constructed before initiating the study and withfeedback from all primary investigators and surgeons based on theircompiled experience with commonly encountered and expected clinicalmanifestations after standard PRK surgery.

One month indicates resolution by the EOS visit, or within 37 dayspost-surgery in cases where an EOS visit is not performed or there ismore than one week between study surgeries.

Adverse events and serious adverse events included an assessment of bothsystemic and ocular assessments. Systemic safety assessments includedregular assessments of height and weight, vital signs, reportingmedication errors including misuse/abuse, pregnancy reporting and earlyphase safety monitoring. The misuse/abuse, pregnancy reporting and earlyphase safety monitoring were not assessed for the study. Ocular safetyassessments included the following:

-   -   Best-corrected visual acuity (BCVA) and uncorrected visual        acuity (UCVA): Was measured at each visit using an ETDRS visual        acuity chart at 4 meters (13 feet) or 1 meter (for patients that        cannot read the 4 meter chart). The BCVA scoring was done based        on the number of correctly read letters plus 30. If visual        acuity was so poor that the patient could not read any of the        largest letters at 1 meter, count fingers and hand movement        vision and light perception was tested.    -   Intraocular Pressure (IOP): IOP measurements were conducted with        applanation tonometry or Tonopen.    -   Dilated fundus exam: The dilated fundus examination included        ophthalmoscopic assessments of the vitreous,        retina/macula/choroid, and optic nerve. Evaluations for retinal        tear/detachment, retinal hemorrhage, vitreous hemorrhage        density, vitreous haze grading and abnormal findings were done        and scored according to grading criteria.    -   Ocular hyperemia: Conjunctival redness of the bulbar conjunctiva        in each eye at the slit lamp according to the McMonnies redness        scale was graded. Hyperemia was assessed in four regions        (superior, inferior, temporal, nasal) of each eye with severity        scored 0-5 in each region.    -   Size of epithelial defect by slit lamp exam: Using an oblique        viewing angle relative to the source beam and a narrow slit        beam, the cornea was sectioned to visualize the vertical and        horizontal borders of the corneal wound. By adjusting the        calibrated slit beam width and height, the maximum horizontal        (wound width) and vertical dimensions (wound height) of the        surgical epithelial wound was estimated. The evaluation was        performed until wound closure and report of wound size as 0        horizontal and 0 vertical dimensions.    -   Slit-lamp biomicroscopy: Ocular signs (eyelids/conjunctiva,        cornea, lens, and iris/anterior chamber) was assessed in both        eyes by slit lamp biomicroscopy according to the grading        criteria.    -   Blink rate: A blink was defined as a bilateral paroxysmal        closure of the eyelids (duration <1 second) in the absence of a        provoking external stimulus. Each blink assessment lasted for 2        min (or as close to 2 minutes as possible with minimum time 1        minute) and the obtained blink rate was averaged to calculate        blink rate in blinks/min.    -   Tear production (Schirmer's test without anesthesia): The test        was performed without anesthetic in both eyes simultaneously.        Tear secretion was measured in millimeters of the length of        strip wetted by tears. The measurement was made to the nearest        whole number.    -   Corneal staining: This test was performed by gently touching the        wet end of an impregnated sodium fluorescein strip to the        inferior conjunctival sac. The strip was wetted with 1 drop of        sterile saline and flicked to remove excess saline. The patient        blinked several times to ensure dispersion of the dye throughout        the tear film prior to grading on a scale of 0-3 (0=Normal, No        staining; 1=Mild, Superficial stippling micropunctate staining;        2=Moderate, Macropunctate staining with some coalescent areas        and 3=Severe, Numerous coalescent macropunctate areas and/or        patches) for each of 5 zones (central plus 4 quadrants).

Safety Evaluation

There were no deaths or SAEs or discontinuation of the drug ordiscontinuation from the study due to AEs in the study. A total of 18AEs were reported in 10 patients (25% of the 40 enrolled patients) ofwhich twelve occurred after treatment with compound I, five aftertreatment with Vehicle and one occurred in one patient prior to dosingof the study drug. All AEs were either mild or moderate in severity.

A total of 10 patients (25%) experienced at least one treatment emergentAE, five patients experienced a single AE, and five patients experiencedmore than one AE. Five patients experienced an AE only during treatmentwith compound I, two patients experienced an AE only during treatmentwith Vehicle and three patients experienced an AE during treatment withboth compound I and Vehicle. There was one AE (headache) that occurredprior to dosing of first study drug.

There were six ocular AEs in four patients (10% of the 40 enrolledpatients), all of which were of mild severity (three eyes each treatedwith compound I and Vehicle). None of the AEs were suspected to berelated to either study drug (compound I or Vehicle). Five of the sixocular AEs were thought to be related to the PRK procedure (two belongedto compound I treated eyes and 3 Vehicle treated eyes) by theInvestigator.

There were twelve non-ocular AEs in seven patients (17.5%) (six eyestreated with compound I, two eyes treated with Vehicle and one patientduring no drug period). Five AEs in four patients (10%) were of moderateseverity (four eyes treated with compound I and one eye treated withVehicle). The remaining AEs were of mild severity. None of the AEs weresuspected to be related to compound I by the investigator.

Table 30 provides the overall incidence of adverse events.

TABLE 30 Overall incidence of AEs - number of events and number ofpatients (Safety analysis set) Compound I 2.5% Vehicle No drug* Total N= 40 N = 40 N = 40 N = 40 n (%) n (%) n (%) n (%) Patients with at leastone AE 8 (20)  5 (12.5) 1 (2.5) 10 (25)  Number of AEs 12  5 1 18 Patients with at least one ocular AE  3 (7.5) 3 (7.5) 0 4 (10) Number ofocular AEs 3 3 0 6 Patients with at least one non-ocular AE 6 (15) 2(5)  1 (2.5)  7 (17.5) Number of non-ocular AEs 9 2 1 12  Number of mildseverity AEs 8 4 1 13  Number of moderate severity AEs 4 1 0 5 Number ofsevere AEs 0 0 0 0 Number of procedure-related AEs 2 3 0 5 Number ofdrug-related AEs 0 0 0 0 An AE starting in one period and continuinginto the next was counted only in the onset period. N = number ofpatients studied. *No drug, or baseline-emergent AE: AEs encounteredafter singing the informed consent and before the administration of anystudy drug (compound I or Vehicle).

Ocular Safety Assessments

Best-corrected visual acuity (BCVA) and uncorrected visual acuity(UCVA): The administration of compound I did not result in any trend orobvious difference between compound I and Vehicle-treated eyesthroughout the study. The data therefore indicate that compound Ireduces pain without adversely affecting BCVA and UCVA.

Intraocular pressure (IOP): No trend across the scheduled time points orobvious differences between compound I and Vehicle-treated eyes wasobserved throughout the study. At the end of study, there was slightincrease (<5 mmHg) in mean IOP values compared to Baseline. The smallchange in mean scores during the study was not clinically significant.

Dilated fundus exam: No abnormal findings were reported by theInvestigator across the scheduled time points for compound I andVehicle-treated eyes.

Ocular hyperemia: A bar chart of ocular hyperemia over time for compoundI and placebo treated eyes is provided in FIG. 8A and FIG. 8B,respectively. FIGS. 8A and 8B indicate the Grade 4 and Grade 3 ocularhyperemia (as measured on the McMonnies scale) on Days 3 and 4,respectively. As seen in FIG. 8A, on Day 2 postoperatively (24 hoursafter PRK surgery) there was less grade 4 hyperemia (all quadrants) incompound I treated eyes compared to Vehicle treated eyes. In thesuperior quadrant the p-value for this difference was 0.04. On Day 3 (48hours post-operative) less grade 3 hyperemia was observed in compound Itreated eyes compared to Vehicle treated eyes. None of the observationswere captured as AEs.

Size of epithelial defect by slit lamp exam: In order to assess the rateof wound healing after administration of compound I, the size of theepithelial defect in both compound I and Vehicle treated populations wasmeasured. The elliptical area of epithelial wound size was calculated asfollows: Area (mm2)=width×height×pi, from the width and height of theepithelial defect measured at the slit lamp. The difference inepithelial wound area between compound I and Vehicle was not noticeablydifferent at any time point except at Day 2 post-PRK surgery (allp-values for the difference in area was >0.35). On Day 2 (24 hourspost-operative), the difference in average epithelial defect areabetween patients treated with compound I vs. Vehicle was 11.23 mm² (pvalue=0.034). This difference was not clinically significant in theimmediate post-operative period. By Day 3 (48 hours post-operative)there was no difference between the compound I-treated and theVehicle-treated eyes and the area of the wound was very small. On Day 4(48 hours post-operative) almost all eyes had healed and there was nodifference between compound I-treated and Vehicle treated eyes. CompoundI showed no delay in wound healing compared to Vehicle.

FIG. 9 and Table 31 provide a comparison of the epithelial size defectof compound I treated eyes versus Vehicle treated eyes.

TABLE 31 Statistical analysis of epithelial defect size (Safety analysisset) Parameter (unit): Epithelial Wound Size (mm) Subcategory: Area(mm²) Raw mean (SD) Difference compound I − Visit compound I VehicleVehicle p-value* Day 1 220.15 (32.26)  217.16 (34.72)   2.98 (27.33)0.494 Day 2 83.02 (32.08) 71.79 (26.96) 11.23 (32.30) 0.034 Day 3  6.75(12.66) 5.73 (9.73) 1.02 (8.78) 0.467 Day 4 0.39 (2.04) 0.08 (0.50) 0.31(2.11) 0.352 Day 8 0.00 (0.00) 0.00 (0.00) 0.00 (0.00) *Obtained from apaired t-test Epithelial defect size was calculated by slit lamp exam asthe area of an ellipse calculated by maximum horizontal and verticaldistance from center of epithelial defect. Area (nun²) = width*height*pi

Slit-lamp biomicroscopy: The slit-lamp biomicroscopic examinationconsisted of examination of ocular structures (eyelids/conjunctiva,cornea, iris/anterior chamber, lens, aqueous flare and aqueousinflammatory cell grade). No abnormalities were observed in ocularstructures of aqueous flare and aqueous inflammatory cell grade,iris/anterior chamber and lens in patients treated with both compound Iand Vehicle across the scheduled time points.

Blink rate: There were fluctuations in the mean blink rate; however, notrend across the scheduled time points or obvious differences betweenblink rate after compound I and Vehicle treatment was observedthroughout the study. At the end of study, no clinically relevantchanges were observed in blink rate compared to Baseline.

Corneal staining: Most of the patients had normal (grade 0) degree ofstaining measured on Baseline, Day 8 and EOS visits. No clinicaldifferences were observed in the corneal staining between the eyestreated with compound I versus Vehicle, at either the baseline, or theDay 8 or the EOS visit.

Vital signs, physical findings and other observations related to safety:The vital sign parameters (systolic and diastolic blood pressures, pulserate and body temperature) were within the normal range for all patientsduring the study. No AEs related to vital signs were observed.Electrocardiogram and special safety topics were not conducted andassessed (as per protocol).

Summary of Safety Results

-   -   There were no deaths, or serious or severe AEs, treatment        discontinuations or study discontinuations reported in this        study.    -   No AEs were evaluated to be related to study drug (either        compound I or Vehicle). Eight of 40 patients and five of 40        patients developed AEs after compound I and Vehicle treatment,        respectively. All Ocular AEs were mild and balanced between        compound I and Vehicle with the vast majority being well known        AEs related to the PRK procedure.    -   There were no observed clinically meaningful differences in        safety between compound I and Vehicle.    -   Compound I showed no delay in wound healing compared to Vehicle.    -   Fewer compound I-treated eyes showed severe conjunctival        hyperemia on Day 2 (24 hours post-operative) compared to        Vehicle-treated eyes.    -   There were no clinically relevant changes observed for BCVA,        IOP, slit-lamp biomicroscopy, dilated eye exam, blink rate, tear        production, corneal staining, or vital signs after compound I        administration compared to Vehicle.

The incidence of AEs by preferred term is presented in Table 32.

TABLE 32 Incidence of AEs by preferred term - n (percent) of patients(Safety analysis set) Compound I 2.5% Vehicle No drug (prior to drug)* N= 40 N = 40 N = 40 n (%) n (%) n (%) Total Procedure Procedure ProcedureN = 40 AEs by preferred term n (%) Severity related n (%) Severityrelated n (%) Severity related n (%) Patients with at least one AE  8(20.0)  5 (12.5) 1 (2.5) 10 (25)   Ocular Corneal Infiltrate 0 1 (2.5)mild yes 0 1 (2.5) AEs Corneal Opacity 1 (2.5) mild yes 1 (2.5) mild yes0 1 (2.5) Punctate Keratitis 1 (2.5) mild yes 1 (2.5) mild yes 0 1 (2.5)Posterior Vitreous 1 (2.5) mild no 0 0 1 (2.5) Detachment Non- Headache1 (2.5) mild no 1 (2.5) moderate no 1 (2.5) mild no 3 (7.5) ocularArthralgia 1 (2.5) mild no 0 0 1 (2.5) AEs Nasopharyngitis 2 (5.0) mild,mild no 0 0 2 (5.0) Oropharyngeal Pain 1 (2.5) mild no 0 0 1 (2.5)Tinnitus 0 1 (2.5) mild no 0 1 (2.5) Sinus Congestion 1 (2.5) moderateno 0 0 1 (2.5) Sinusitis 1 (2.5) moderate no 0 0 1 (2.5) Pyrexia 1 (2.5)moderate no 0 0 1 (2.5) Vomiting 1 (2.5) moderate no 0 0 1 (2.5) Anadverse event starting in one period and continuing into the next iscounted only in the onset period. N = number of patients studied. n =number of patients with at least one AE in the category. *No drug, orbaseline-emergent AE: AEs encountered after singing the informed consentand before the administration of any study dmg (compound I or Vehicle).

All publications and patent documents cited herein are incorporatedherein by reference as if each such publication or document wasspecifically and individually indicated to be incorporated herein byreference. The present invention and its embodiments have been describedin detail. However, the scope of the present invention is not intendedto be limited to the particular embodiments of any process, manufacture,composition of matter, compounds, means, methods, and/or steps describedin the specification. Various modifications, substitutions, andvariations can be made to the disclosed material without departing fromthe spirit and/or essential characteristics of the present invention.Accordingly, one of ordinary skill in the art will readily appreciatefrom the invention that later modifications, substitutions, and/orvariations performing substantially the same function or achievingsubstantially the same result as embodiments described herein may beutilized according to such related embodiments of the present invention.Thus, the following claims are intended to encompass within their scopemodifications, substitutions, and variations to processes, manufactures,compositions of matter, compounds, means, methods, and/or stepsdisclosed herein. The claims should not be read as limited to thedescribed order or elements unless stated to that effect. It should beunderstood that various changes in form and detail may be made withoutdeparting from the scope of the appended claims.

1-94. (canceled)
 95. A method of treating ocular surface pain in asubject in need thereof, comprising ocularly administering to thesubject a formulation, comprising: a suspension of4-(7-Hydroxy-2-isopropyl-4-oxo-4H-quinazolin-3-yl)-benzonitrile(compound I) or a salt, co-crystal, or polymorph thereof, in an amountof about 0.5% w/v to about 3.5% w/v, tyloxapol in an amount of about0.04% w/v to about 0.06% w/v; carbomer homopolymer Type B in an amountfrom about 0.05% w/v to about 0.4% w/v; a tonicity agent; a buffer;sodium chloride in an amount of from 0.01% w/v to about 1% w/v;optionally, a preservative; and water in quantity sufficient (qs) to100%.
 96. The method according to claim 95, wherein the tonicity agentis a polyol selected from the group selected from mannitol, glycerin,xylitol, sorbitol, propylene glycol, and combinations thereof.
 97. Themethod according to claim 95, wherein the buffer is selected from thegroup consisting of acetate, ascorbate, borate, hydrogen carbonate,carbonate, citrate, edetate (EDTA) gluconate, lactate, phosphate,propionate and TRIS (tromethamine).
 98. The method according to claim95, wherein the pH of the formulation is about 5.0 to about 8.0, about5.5 to about 8.0, about 5.5 to about 7.5, about 5.0 to about 7.4, about5.5 to about 7.4, about 6.0 to about 8.0, about 6.5 to about 8.0, about6.0 to about 7.4, or about 6.5 to about 7.4.
 99. The method according toclaim 95, further comprising an additional agent selected from the groupconsisting of polyvinylprrolidone, hydroxypropylβcyclodextrin, andsulfoalkylether β-cyclodextrin in an amount of at least about 1.5 w/v %,at least about 3.0 w/v %, at least about 3.5 w/v % or at least about 4.5w/v, but no greater than about 10.0 w/v %, no greater than about 8.0%w/v, no greater than about 6.5 w/v %, or no greater than about 5.5 w/v.100. The method according to claim 95, further comprising an additionalagent selected from the group consisting of cyclodextrins in an amountof at least about 1.5 w/v %, at least about 3.0 w/v %, at least about3.5 w/v % or at least about 4.5 w/v, but no greater than about 10.0 w/v%, no greater than about 8.0% w/v, no greater than about 6.5 w/v %, orno greater than about 5.5 w/v.
 101. The method according to claim 95,wherein compound I is in polymorphic form B.
 102. The method accordingto claim 101, wherein the polymorphic form B is characterized by anX-ray diffraction pattern having three or more peaks at 2θ valuesselected from 9.3, 10.6 and 14.4.+−.0.2° 2θ.
 103. The method accordingto 95, wherein the subject suffers from one or more of dry eye disease,Sjogren's Syndrome, conjunctivitis (including keratoconjuctivitis,vernal keratoconjunctivitis, allergic conjunctivitis),Map-Dot-Fingerprint Dystrophy, acanthamoeba, fibromyalgia, Meibomiangland dysfunction, thyroid eye disease, rosacea, ptosis, keratoconus,ocular pain syndrome, Steven-Johnson's syndrome, cornealepitheliopathies, corneal neuropathies (including LASIK induced cornealneuropathies), corneal dystrophies (including recurrent cornealdystrophies), epithelial basement membrane dystrophy, corneal erosionsor abrasions (including recurrent corneal erosions or abrasions), ocularsurface diseases, blepharitis, graft vs host disease, meibomitis,glaucoma, conjunctivochalasis, keratopathis (including herpetickeratopathy, filamentary keratopathy, band or bullous keratopathy,exposure keratopathy), keratitis (including herpes simplex viruskeratitis), iritis, episclentis, corneal surgery, multiple sclerosis,trichiasis, pterygium, neuralgia, xerophthalmia, patients recoveringfrom neurotrophic keratitis, or ocular pain persisting for at leastthree months after photorefractive keratectomy (PRK) surgery orlaser-assisted in situ keratomileusis (LASIK) surgery.